Dose-dependent effects of cadmium on food consumption, mass, and bioaccumulation in the moth Spodoptera littoralis

Dose-dependent effects of cadmium on food consumption, mass, and bioaccumulation in the moth Spodoptera littoralis | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Dose-dependent effects of cadmium on food consumption, mass, and bioaccumulation in the moth Spodoptera littoralis Ségolène Humann-Guilleminot, Marie-Anne Pottier, Maryse Rouelle, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5439205/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Our environment is increasingly contaminated by various substances, mainly from industrial and agricultural activities. Among the main pollutants, heavy metals, are highly toxic to a wide range of organisms including invertebrates. In our study, we focused on the impacts of cadmium on the model pest moth Spodoptera littoralis . In laboratory conditions, larvae were exposed to a diet contaminated with nine different cadmium concentrations ranging from 7.80 µg/g to 50 mg/g of food, over six developmental stages. Our results show that sublethal doses of cadmium have a significant effect on food consumption and relative larval growth rate over a 6-day period, and to a lesser extent, affect larval mass at higher doses. Adverse effects were noted at a sublethal dose of 31.25 µg/g, with particularly severe outcomes at the highest lethal doses ranging from 0.5 to 50 mg/g, where 100% mortality was observed after treatment. While higher doses from 250 µg/g influenced larval mass, most larvae recovered and regained mass, except for those exposed to the two highest doses just prior metamorphosis. In addition, our results indicated that cadmium accumulation reached its maximum at pupal stage and decreased in adults, suggesting that efficient detoxification processes occur during metamorphosis, with significant amounts of cadmium that were excreted through the integument and exuviae at the pre-pupal stage. Moreover, moths that ingested cadmium at larval stage have stored most of it in their digestive tract, from where it has been transferred to various parts of the body, including the head and antennae, via haemolymph. Cadmium Heavy metal Moth Growth Bioaccumulation Insect Figures Figure 1 Figure 2 Figure 3 1. Introduction In recent decades, the accumulation of environmental pollutants in various ecosystems, largely due to industrial and agricultural activities, has raised growing concern (Gao and Wen, 2016 ; Jin et al., 2020 ; Nagorka and Koschorreck, 2020 ; Vardhan et al., 2019 ). Among pollutants, heavy metals accumulate at varying concentrations in the environment and in animals, resulting in a wide range of physiological effects (Ali and Khan, 2018 ; Tabassum et al., 2024 ; Verma et al., 2023 ). Due to their toxicity, persistence and bioaccumulation capacity, they pose significant long-term risks as they can remain in ecosystems for centuries (Ali et al., 2019 ; Brunn et al., 2023 ; Net et al., 2015 ). Cadmium, a heavy metal element, is released into the environment from various sources including industrial wastes and agricultural runoff (Bradl, 2005 , Kubier et al., 2019 ). Like other metals, cadmium is persistent in the environment and accumulates in living organisms, with potential adverse effects on growth and acting as an endocrine disruptor (Ali and Khan, 2018 ; Raina et al., 2001 ; Sorvari et al., 2007 ; Verma et al., 2023 ). The toxic effects of cadmium are well-documented and affect various physiological processes in microorganisms, plants and animals. However, its impact varies by species and developmental stages, with some organisms showing a higher propensity to bioaccumulate and adverse effects (Vardhan et al., 2019 ). For instance, cadmium is known to alter the expression of hormone receptor genes and to disrupt circulating hormone levels, which has negative effects on moulting, growth and reproduction in various invertebrate species (Iavicoli et al., 2009 ; Planelló et al., 2010 ; Rana, 2014 ). In several insect species, such as the flies Boettcherisca peregrina, Drosophila melanogaster and Musca domestica , high doses of cadmium ingestion delay the transitions from larval to adult development, and similar findings were reported for mercury (Frat et al., 2021 ; Raina et al., 2001 ; Wu et al., 2006 ). In addition, studies have shown a detrimental negative impact of cadmium and other heavy metals on mass and survival of blow flies, particularly in larval stages (review in Raise and Gemmellaro, 2024 ). In a previous study on the model pest moth Spodoptera littoralis , we showed that ingestion of cadmium at low and high (sublethal) doses delayed post-embryonic development and affected larval growth and survival (Humann-Guilleminot et al., 2024 ). As a widespread agricultural pest, S. littoralis interacts closely with various ecosystems and is frequently exposed to different pollutants (EFSA, 2015). Its short life cycle and well-documented developmental stages make it an ideal model for studying the effects of pollutants over a full life cycle (Brown and Dewhurst, 1975 ). In addition, its physiological and developmental processes are similar to those of other insects, allowing the findings to be extrapolated into a broader ecological context. To better understand the toxicity of cadmium in this species, we experimentally evaluated the effects on dietary intake of nine doses of cadmium, ranging from sublethal to lethal, administered orally to larvae, on their food consumption, mass gain, relative growth rate, and mortality over a 6-day period. We also examined the bioaccumulation of two sublethal doses of ingested cadmium at different developmental stages, from the fourth larval stage to adulthood. We studied the distribution of cadmium in the tissues of final larval and pre-pupal stages, and measured its concentrations in tissues and haemolymph of adults. We hypothesized that cadmium would have a negative impact on all measured variables, including food intake, mass gain, relative growth rate and larval mortality, with more pronounced effects at higher doses. In addition, we expected that cadmium would be detected in all individuals that ingested it, with higher concentrations corresponding to higher doses. We also predicted an increase in the concentration of cadmium at the larval stage, followed by a decrease at adulthood, due to detoxification during metamorphosis. 2. Material and methods 2.1. Moths rearing and exposure to treatment S. littoralis larvae were reared on a semi-artificial diet, as described in (Humann-Guilleminot et al., 2024 ). They were stored in glass jars at 4°C until use, in order to ensured minimal chemical contamination, as both treated and control diets were stored in glass rather than plastic containers. Larvae were raised in DEHP-free plastic boxes, which were replaced regularly for each experiment. All laboratory materials have been sterilized to avoid microbial contamination and excessive mortality. The uniform distribution and absorption of cadmium in the diet was achieved by incorporating cadmium (CdCl 2٠ 2.5H 2 O, 239208-100G, Sigma, France) into food that was in liquid form. The mixture has been mixed with an industrial stirrer for an extended period. Agar was included to solidify the mixture into a uniform jelly after cooling and then consumed by insect larvae. From the fourth larval instar to pupation (corresponding to about 15 days), larvae were fed with food containing nine cadmium concentrations (“T1” to “T9”) prepared from a 3M solution, i.e. 7.8, 15.625, 31.25, 62.5, 125, 250, 500 µg, 5 and 50 mg CdCl 2 per gram of food. Control larvae received a diet mixed with solvent solution (500 µL of ethanol in 100 g semi-solid food). Larvae were fed individually ad-libitum under specific contamination conditions. 2.2. Assessment of larval food consumption, growth and mortality The experimental phase included 200 larvae (N=20 per condition). The food was weighed daily to calculate the rate of consumption according to the CdCl 2 concentration, and the larvae were also weighed daily until pupation. The relative growth rate was calculated using the formula from Farrar et al., 1989: (average mass at day 1 and day 6). Finally, the mortality rates at the larval and pupal stages were recorded for each treatment. 2.3. Cadmium quantification To evaluate cadmium bioaccumulation, we quantified cadmium in treated and control insects using an atomic absorption spectrometer. Samples were analysed after a mineralization process in a Teflon flask at 100°C. They were mixed with 1 ml of 68% nitric acid (HNO₃; VWR Analar Normapur) for 24 hours, then with 0.5 ml of 30% hydrogen peroxide (H₂O₂; VWR Analar Normapur) for again 24 hours. Acid mineralization was performed to break down the sample matrix, releasing the elements into a solution for further analysis. Once mineralized, the liquid samples were transferred into tubes (215–0329 from VWR) and diluted with 6 ml of MilliQ water (Purelab Ultra system from Elga). The weight of each tube was recorded for precise measurement of its content. Depending on the analyses, the insects were fed different doses of cadmium and different tissues, or body parts were dissected to perform the measurements. To follow cadmium concentrations over postembryonic development, two groups of larvae received either 15.625 (T2) or 125 µg (T5) of CdCl 2 from the 4th larval stage to adulthood. At least three insects were sampled daily for whole-body analysis. Larvae were fasted for 24 hours to clean their digestive tracts before any measurements. To evaluate cadmium concentration in different tissues at various stages of development, including final larval, pre-pupal and adult stages, two other larval groups were fed either 62.5 (T4) or 125 µg (T5) cadmium using set of individuals that differed from those used for whole-body measurements. For the final larval stage, we analysed the digestive tract, the fat body and the head. For the pre-pupal stage, cadmium levels were measured in the feces, in exuviae and integument. In male and female adults, measurements were made from the head, thorax, abdomen, antennae and haemolymph. All tissues were thoroughly rinsed in Ringer’s solution, especially the digestive tract. Haemolymph was collected from one-day-old adults, while larval feces were collected throughout the study. For each stage and each treatment, we analysed five samples in each category. To ensure sufficient analysis, tissues were pooled in triplicate to achieve a minimum of 50 mg of material, then dried at 150°C for 10 to 15 minutes. Exceptions to this drying protocol were haemolymph, antennae, and exuviae, which were handled differently. We then measured the quantity of cadmium in each pool using Inductively Coupled Plasma Optical Emission Spectrometry (ICP-OES Agilent 5100SDVD, Agilent Technologies, Santa Clara, CA, U.S.A.) at the ALIPP6 platform (Sorbonne University, Paris, France), with a detection threshold of ICP-OES at 3 mg/kg. 2.4. Statistical analyses All statistical analyses were performed using the R v. 4.2.2 software (R Core Team, 2022 ). For each analysis, the control group was set as the reference using the relevel function from the stats package. 2.4.1. Effect of cadmium on larval food consumption, mass, relative growth rate and mortality We first aimed to assess the evolution of both larval food consumption and mass from the 4th larval stage to pre-pupae under ten different treatments (9 doses of cadmium and control). We ran two linear mixed effect models using the lmer function from the R package lmerTest (Kuznetsova et al., 2017 ), that included either the logit-transformed percentage of food consumption or log-transformed body mass as the response variable, day (from 0 to 6, continuous), treatment (categorical), interaction between day and treatment, and quadratic term of day and interaction between quadratic term day and treatment to account for the observed non-linear relationship between time and food consumption, or mass, at larval stage day 0 relative to larval stage at day 6 as explanatory variables. The model included individual identity as random intercept to account for the repeated measures taken within each individual. We then conducted a linear model using the lm from the stats package to assess the influence of cadmium treatments on larvae relative growth rates, that included the relative growth rate as the response variable and the treatment as the explanatory variable. Lastly, we conducted a generalized linear model with a binomial distribution to assess the effect of treatments on larval mortality. The model was implemented using the glm function from the stats package. The response variable was the binary outcome of larval mortality, with treatment as the explanatory variable. However, since larval mortality was 100% in the treatments with the three highest cadmium doses (T7, T8, T9), these treatments were excluded from the analysis. 2.4.2. Bioaccumulation of cadmium across developmental stages and tissues We analysed the differences in bioaccumulation of two sublethal doses (T2 and T5) of ingested cadmium at different developmental stages and in various tissues. We first applied a linear model on a first set of data using the lm function from the stats package, with log-transformed cadmium concentration as the response variable, and stage (from the 4th to the 6th, pre-pupae, 3-day pupae, 9-day pupae, and adults), treatment, and the interaction between stage and treatment as explanatory variables. Furthermore, we investigated cadmium concentrations in different tissues extracted from individuals at three different stages of development. We thus applied three linear models at larval, pre-pupal and adult stages using the lm function from the stats package, with either untransformed or square-root transformed cadmium concentration as the response variable, the tissue, the treatment, and the interaction between tissue and treatment as explanatory variables. We then conducted and analysis of variance using the Anova function from the car package. Post-hoc tests analyses were performed using the lsmeans function from the emmeans package. As there was no evidence of a difference in cadmium concentration between males and females at any stage (all p > 0.10), the sex of individuals was not included in the analysis. For linear-mixed effect model with interaction terms, omnibus analyses of variance were conducted using the Kenward-Roger approximation for the computation of fixed effects degrees of freedom using the R package lmerTest (Kuznetsova et al., 2017 ). Modelling assumptions (normality of residuals, normality of random effects, residuals linearity, homogeneity of variance, collinearity of factors) were checked by visual inspection of the residuals using the check_model function of the performance package (Lüdecke et al., 2021 ). Following recommendations of the American Statistical Association (Wasserstein and Lazar, 2016 ), we do not use an arbitrary threshold (e.g. 0.05) to interpret our results, but instead we use p-values to assess the strength of the statistical evidence to reject the null hypothesis. 3. Results 3.1. Effect of treatment on food consumption, body mass, relative growth rate and larval mortality Summary statistics (sample size, median, mean ± sd, min and maximum values) for food consumption, mass and relative growth rate across developmental stages and treatments are presented in Table S1 – S3. To explore the relationship between cadmium treatments and individual food consumption or larval mass, our models were designed to compare changes in these variables over six days for each treatment. The inclusion of a quadratic term proved to be the most suitable approach, particularly due to the distribution of the data and marked changes in food consumption or mass observed between day 0 and day 6. Food intake on day 0 was 100% for all treatments and there was no difference in mass at day 0 between cadmium the treated subjects and the controls (p = 0.71). In addition, the food consumption of larvae from the highest treatment T9 on day 6 could not be monitored as all larvae died before data collection. We observed small to strong statistical evidence suggesting that larvae from treatments T3 (31.25 µg) and from T5 (125 µg) to T9 (50 mg) exhibited higher food consumption on day 1 compared to the control group (Table 1 , Fig. 1 ). In addition, T3 to T9 treatments, including T4 (62.50 µg), showed a distinct pattern from day 1 to day 4, with food consumption consistently higher than that of the control group. From day 4 to day 6, this pattern of increased food consumption became even more pronounced in these treatments than in the controls. This trend correlates with the interaction between day and treatment and the linear component of the quadratic term model (Table 1 , Fig. 1 ). Table 1 Summary of the linear mixed model investigating the effect of treatments on food consumption (logit-transformed) over six days, fitted using a restricted maximum likelihood function. Model includes continuous variable days, treatment, the interaction between days and treatment as well as quadratic terms. The table shows model estimates ± standard error (Est. ± s.e.), associated 95% confidence intervals (C.I.) and p-values. Fixed effects Est. ± s.e. C.I. P Intercept (day 1, control) 3.45 ± 0.04 [3.37–3.53] < 0.001 Day 0.07 ± 0.03 [0.00–0.13] 0.034 Treatment T1–7.80 µg 0 ± 0.06 [-0.11–0.11] 0.969 T2–15.625 µg -0.03 ± 0.06 [-0.14–0.08] 0.57 T3–31.25 µg 0.11 ± 0.06 [0.00–0.22] 0.045 T4–62.50 µg 0.09 ± 0.06 [-0.02–0.20] 0.102 T5–125 µg 0.13 ± 0.06 [0.02–0.24] 0.025 T6–250 µg 0.14 ± 0.06 [0.03–0.25] 0.015 T7–0.5 mg 0.17 ± 0.06 [0.06–0.29] 0.002 T8–5 mg 0.15 ± 0.06 [0.04–0.26] 0.009 T9–50 mg 0.19 ± 0.06 [0.07–0.30] 0.002 Day 2 -0.04 ± 0 [-0.05 – -0.03] < 0.001 Day × Treatment T1–7.80 µg -0.03 ± 0.04 [-0.12–0.05] 0.465 T2–15.625 µg 0.02 ± 0.04 [-0.07–0.11] 0.652 T3–31.25 µg -0.09 ± 0.04 [-0.18 – -0.01] 0.032 T4–62.50 µg -0.09 ± 0.04 [-0.18 – -0.01] 0.034 T5–125 µg -0.09 ± 0.04 [-0.17 – -0.00] 0.048 T6–250 µg -0.12 ± 0.04 [-0.20 – -0.03] 0.008 T7–0.5 mg -0.2 ± 0.04 [-0.29 – -0.12] < 0.001 T8–5 mg -0.07 ± 0.04 [-0.16–0.01] 0.092 T9–50 mg -0.12 ± 0.06 [-0.24 – -0.00] 0.044 Day 2 × Treatment T1–7.80 µg 0.01 ± 0.01 [-0.00–0.03] 0.096 T2–15.625 µg 0 ± 0.01 [-0.01–0.01] 0.962 T3–31.25 µg 0.01 ± 0.01 [0.00–0.03] 0.037 T4–62.50 µg 0.02 ± 0.01 [0.00–0.03] 0.017 T5–125 µg 0.02 ± 0.01 [0.00–0.03] 0.008 T6–250 µg 0.03 ± 0.01 [0.02–0.05] < 0.001 T7–0.5 mg 0.06 ± 0.01 [0.05–0.08] < 0.001 T8–5 mg 0.03 ± 0.01 [0.02–0.05] < 0.001 T9–50 mg 0.05 ± 0.01 [0.02–0.07] < 0.001 Similarly, the difference in mass between the larvae treated with higher doses (T6 to T9) and the control group increased over time. Larvae from treatments T6 and T7 showed a lower mass on day 1 compared to the control group, with this difference decreasing over time (i.e. individuals in this group compensate for their delayed mass gain compared to the control group). However, for treatments T8 and T9, the mass remains lower throughout the study period, indicating a sustained negative impact of these highest doses on larval mass (Table 2 , Fig. 1 ). Table 2 Summary of the linear mixed model investigating the effect of treatment on mass (log-transformed) over six days fitted using a restricted maximum likelihood function. Model includes continuous variable days, treatment, the interaction between days and treatment as well as quadratic terms. The table shows model estimates ± standard error (Est. ± s.e.) and associated 95% confidence intervals (C.I.); for log-transformed variables, the table also shows multiplicative estimates for untransformed variable (Est. mult.) and associated multiplicative confidence intervals (C.I. mult.); p-values. The multiplicative estimate of the intercept corresponds to the geometric mean of the variable. Fixed effects Est. ± s.e. C.I. Est. Mult. C.I. mult. P Intercept (day 0, control) -3.12 ± 0.07 [-3.25 – -2.99] 0.04 [0.04–0.05] < 0.001 Day 0.56 ± 0.05 [0.46–0.66] 1.75 ± 0.09 [1.59–1.93] < 0.001 Treatment T1–7.80 µg 0 ± 0.09 [-0.18–0.17] 1 ± 0.09 [0.83–1.19] 0.957 T2–15.625 µg -0.02 ± 0.09 [-0.20–0.16] 0.98 ± 0.09 [0.82–1.17] 0.798 T3–31.25 µg -0.07 ± 0.09 [-0.25–0.11] 0.93 ± 0.08 [0.78–1.11] 0.424 T4–62.50 µg -0.07 ± 0.09 [-0.25–0.11] 0.93 ± 0.08 [0.78–1.12] 0.454 T5–125 µg -0.14 ± 0.09 [-0.32–0.04] 0.87 ± 0.08 [0.73–1.04] 0.132 T6–250 µg -0.06 ± 0.09 [-0.24–0.12] 0.94 ± 0.09 [0.79–1.12] 0.505 T7–0.5 mg -0.07 ± 0.09 [-0.25–0.11] 0.93 ± 0.08 [0.78–1.11] 0.422 T8–5 mg 0.07 ± 0.09 [-0.11–0.25] 1.08 ± 0.1 [0.90–1.29] 0.425 T9–50 mg 0.09 ± 0.09 [-0.09–0.27] 1.09 ± 0.1 [0.91–1.31] 0.329 Day 2 -0.01 ± 0.01 [-0.03–0.00] 0.99 ± 0.01 [0.97–1.00] 0.096 Day × Treatment T1–7.80 µg 0.05 ± 0.07 [-0.09–0.19] 1.05 ± 0.07 [0.91–1.21] 0.501 T2–15.625 µg 0.01 ± 0.07 [-0.13–0.15] 1.01 ± 0.07 [0.88–1.16] 0.873 T3–31.25 µg 0.02 ± 0.07 [-0.12–0.16] 1.02 ± 0.07 [0.89–1.18] 0.759 T4–62.50 µg -0.05 ± 0.07 [-0.19–0.09] 0.95 ± 0.07 [0.83–1.09] 0.464 T5–125 µg -0.1 ± 0.07 [-0.24–0.04] 0.9 ± 0.06 [0.79–1.04] 0.153 T6–250 µg -0.2 ± 0.07 [-0.34 – -0.06] 0.82 ± 0.06 [0.71–0.94] 0.004 T7–0.5 mg -0.2 ± 0.07 [-0.34 – -0.06] 0.82 ± 0.06 [0.71–0.94] 0.005 T8–5 mg -0.68 ± 0.07 [-0.82 – -0.54] 0.5 ± 0.04 [0.44–0.58] < 0.001 T9–50 mg -0.71 ± 0.07 [-0.85 – -0.57] 0.49 ± 0.03 [0.43–0.56] < 0.001 Day 2 × Treatment T1–7.80 µg -0.01 ± 0.01 [-0.03–0.01] 0.99 ± 0.01 [0.97–1.01] 0.341 T2–15.625 µg 0 ± 0.01 [-0.02–0.02] 1 ± 0.01 [0.98–1.02] 0.859 T3–31.25 µg 0 ± 0.01 [-0.02–0.02] 1 ± 0.01 [0.98–1.02] 0.954 T4–62.50 µg 0.01 ± 0.01 [-0.01–0.03] 1.01 ± 0.01 [0.99–1.03] 0.419 T5–125 µg 0.01 ± 0.01 [-0.01–0.04] 1.01 ± 0.01 [0.99–1.04] 0.245 T6–250 µg 0.01 ± 0.01 [-0.01–0.04] 1.01 ± 0.01 [0.99–1.04] 0.218 T7–0.5 mg -0.02 ± 0.01 [-0.04–0.00] 0.98 ± 0.01 [0.96–1.00] 0.116 T8–5 mg 0.04 ± 0.01 [0.01–0.06] 1.04 ± 0.01 [1.01–1.06] 0.001 T9–50 mg 0.02 ± 0.01 [0.00–0.05] 1.02 ± 0.01 [1.00–1.05] 0.039 Comparison of the relative growth rate (RGR) between treatments confirms the results previously described, with strong statistical evidence for a decrease in RGR from treatment T5 to T9 (Table 3 , Fig. 1 ). Table 3 Summary of the linear mixed model investigating the effect of treatment on Relative Growth Rate fitted using a restricted maximum likelihood function. Model includes treatment only. The table shows model estimates ± standard error (Est. ± s.e.) and associated 95% confidence intervals (C.I.). Fixed effects Est. ± s.e. C.i. p Intercept (control) 3.37 ± 0.24 [2.90–3.85] < 0.001 Treatment T1–7.80 µg -0.31 ± 0.34 [-0.98–0.37] 0.372 T2–15.625 µg -0.18 ± 0.34 [-0.86–0.49] 0.59 T3–31.25 µg 0.08 ± 0.34 [-0.59–0.76] 0.808 T4–62.50 µg -0.29 ± 0.34 [-0.97–0.38] 0.393 T5–125 µg -0.91 ± 0.34 [-1.59 – -0.24] 0.008 T6–250 µg -2.05 ± 0.34 [-2.72 – -1.38] < 0.001 T7–0.5 mg -3.09 ± 0.34 [-3.77 – -2.42] < 0.001 T8–5 mg -3.33 ± 0.34 [-4.00 – -2.66] < 0.001 T9–50 mg -3.38 ± 0.35 [-4.07 – -2.69] < 0.001 Lastly, the larval mortality of each treatment revealed a lethal dose threshold, since 100% of the larvae died in T7, T8 and T9 treatments, while no difference in mortality rates was found between the other treatments and the control group (all p > 0.10; Fig. 1 ). 3.2. Bioaccumulation of cadmium across developmental stages and tissues For both T2 (15.625 µg) and T5 (125 µg) treatments, cadmium assays demonstrate dose-dependent bioaccumulation during the post-embryonic development compared to the control group (Table 4, Fig. 2 ), with variations according to stages and treatment. Notably, the T2 subjects experienced minor fluctuations compared to those exposed to the highest dose, with a peak at 9-day in the pupal stage. Interestingly, cadmium levels in T5 adults were similar to those of the 4th larval stage, indicating a decrease as the larvae matured into adults (Table 4 and Fig. 2 ). Table 4: Summary of the linear mixed model investigating the effect of Spodoptera littoralis developmental stages and treatment on log-transformed cadmium concentration in individuals using a restricted maximum likelihood function. Model includes developmental stages, treatment and the interaction between stages and treatment. The table shows model estimates ± standard error (Est. ± s.e.) and associated 95% confidence intervals (C.I.); for log-transformed variables, the table also shows multiplicative estimates for untransformed variable (Est. mult.) and associated multiplicative confidence intervals (C.I. mult.); p-values. The multiplicative estimate of the intercept corresponds to the geometric mean of the variable. Fixed effects Est. ± s.e. C.I. Est. Mult. C.I. mult. P Intercept (L4, control) -4.61 ± 0.32 [-5.25 – -3.97] 0.01 [0.01–0.02] < 0.001 Stage L5 1.57 ± 0.38 [0.82–2.32] 4.8 [2.27–10.14] < 0.001 L6 0.05 ± 0.36 [-0.66–0.76] 1.05 [0.52–2.14] 0.89 Pre-pupae 0.07 ± 0.37 [-0.67–0.81] 1.07 [0.51–2.24] 0.85 3-days pupae 0.14 ± 0.40 [-0.65–0.92] 1.15 [0.52–2.50] 0.73 9-days pupae -0.03 ± 0.40 [-0.81–0.76] 0.97 [0.45–2.13] 0.95 Adult 0.01 ± 0.40 [-0.77–0.79] 1.01 [0.46–2.21] 0.98 Treatment 15.625 µg 6.34 ± 0.40 [5.55–7.12] 564.53 [258.39–1233.39] < 0.001 125 µg 8.26 ± 0.40 [7.47–9.04] 3846.81 [1760.70–8404.59] < 0.001 Stage × Treatment Stage L5 × 15.625 µg -1.32 ± 0.48 [-2.27 – -0.37] 0.27 [0.10–0.69] 0.01 Stage L6 × 15.625 µg 0.26 ± 0.46 [-0.65–1.18] 1.3 [0.52–3.25] 0.57 Stage pre-pupae × 15.625 µg 0.80 ± 0.47 [-0.14–1.74] 2.23 [0.87–5.69] 0.09 Stage 3-days pupae × 15.625 µg 1.00 ± 0.51 [-0.01–2.01] 2.72 [0.99–7.47] 0.05 Stage 9-days pupae × 15.625 µg 1.18 ± 0.51 [0.17–2.19] 3.25 [1.18–8.91] 0.02 Stage adult × 15.625 µg 1.01 ± 0.54 [-0.05–2.07] 2.75 [0.95–7.92] 0.06 Stage L5 × 125 µg -1.24 ± 0.46 [-2.15 – -0.32] 0.29 [0.12–0.72] 0.01 Stage L6 × 125 µg -0.43 ± 0.47 [-1.35–0.50] 0.65 [0.26–1.65] 0.36 Stage pre-pupae × 125 µg 0.61 ± 0.50 [-0.39–1.60] 1.84 [0.68–4.97] 0.23 Stage 3-days pupae × 125 µg 0.73 ± 0.50 [-0.25–1.72] 2.09 [0.78–5.58] 0.14 Stage 9-days pupae × 125 µg 0.95 ± 0.51 [-0.06–1.96] 2.6 [0.95–7.12] 0.06 Stage adult × 125 µg 0.22 ± 0.51 [-0.79–1.22] 1.24 [0.45–3.40] 0.67 To go further, cadmium concentrations after T4 and T5 treatments were measured in various tissues of the final larval stage, pre-pupa and adult (Fig. 3 ). We found a difference in cadmium concentration between treatment and the interaction between tissue-treatment for all the three stages (Table 5 ). In the larval control group, a low amount of cadmium was detected in the digestive tract and in the head, likely due to a slight contamination during the measurement process. The other control samples showed levels below the detection limit. Furthermore, post-hoc tests showed no statistical difference in cadmium concentration between the different tissues of the control group at any stage. In contrast, T5-treated larvae exhibited accumulation of cadmium in their digestive tract and fat body. The tissues with the highest bioaccumulation are not the same according to treatment. For example, during the pre-pupal stage, the higher concentrations were observed in the feces of the T5 group but in the exuviae of the T4 (Fig. 3 ). The group exposed to the highest level of cadmium showed the most substantial accumulation in most tissues, except for the integument. In addition, cadmium levels in the feces of pre-pupae reached up to four times the ingested dose and between 10 to 25% of the cadmium was found in the integument, but we did not report any difference in concentration between these two matrices (Fig. 3 ). In T4 and T5-treated adults, cadmium predominantly concentrated in the abdomen (average of 59.98 µg/g and 32.97 µg/g, respectively). Cadmium concentrations were barely detectable in T4-adult antennae and haemolymph (average of 0.40 µg/g and 0.04 µg/g, respectively) and slightly higher in T5 adults (average of 2.53 µg/g and 1.56 µg/g, respectively). Table 5 Analysis of variance testing the difference in cadmium concentrations in different tissues and treatment using Kenward-Roger approximation for the computation of fixed effects degrees of freedom. The table shows the sum of squares, degrees of freedom (Df), F and P values. Larvae Fixed effects Sum of squares Df F P Tissues 17.275 2 75.639 < 0.001 Treatment 248.976 2 1090.171 < 0.001 Tissues × Treatment 50.791 4 111.196 < 0.001 Pre-pupae Fixed effects Sum of squares Df F P Tissues 248.36 2 190.83 < 0.001 Treatment 1546.5 2 1188.26 < 0.001 Tissues × Treatment 347.79 4 133.61 < 0.001 Adults Fixed effects Sum of squares Df F P Tissues 10967.8 4 66.63 < 0.001 Treatment 1985.3 2 24.121 < 0.001 Tissues × Treatment 7074.8 8 21.49 < 0.001 4. Discussion In this study, we explored the impact of ingested cadmium at sublethal to lethal doses on several life history traits of the model crop pest Spodoptera littoralis , including food intake by larvae, post-embryonic development and bioaccumulation. Our results indicate that sublethal doses of cadmium significantly affect food consumption and relative growth rate in larvae over a 6-day period, and to a lesser extent affect their mass after treatments at higher doses. The adverse effects become pronounced at a relatively low-dose of 31.25 µg/g, with particularly severe outcomes at the highest – lethal – doses ranging from 0.5 to 50 mg/g, where 100% mortality was observed post-treatment. Additionally, while higher cadmium treatments starting from 250 µg/g impacted larval mass, we observed that by day 6 before metamorphosis, larvae receiving sublethal doses had regained mass comparable to the control group, suggesting a selective pressure that allows individuals with initially lower mass to compensate for their delayed growth despite reduced food intake. Contrasting findings have been reported in other studies regarding the impact of cadmium on mass, where a decrease in larval body mass was noted under sublethal cadmium exposure, but not always with a recovery to control levels (Humann-Guilleminot et al., 2024 ; Vlahović et al., 2017 , 2014 ). The variation in larval mass may stem from the species' ability to detoxify, a process that can vary in energy consumption (Castañeda et al., 2009 ; Vlahović et al., 2017 ). Specifically, Spodoptera littoralis is believed to possess a high capacity for detoxification (Aviles et al., 2019 ). The dose-dependent relationship observed after the treatments in the present study is consistent with the growth reductions previously reported in this moth (Humann-Guilleminot et al., 2024 ) and in other species belonging to different insects orders (e.g., Aiolopus thalassinus , Oncopeltus fasciatus , Lymantria dispar , Chironomus riparius , Poecilus cupreus ) chronically exposed to heavy metals (Cervera et al., 2004 ; Maryanski et al., 2002 ; Ortel, 1995 ; Schmidt et al., 1992 ; Sildanchandra and Crane, 2000 ). Some studies reported a potential behavioural adaptation of larvae to avoid cadmium contaminated food, where larvae decrease food consumption to ensure survival, thus reducing the intake of cadmium – but also essential nutrients – at the expense of its growth (Migula and Binkowska, 1993 ; van Straalen and Hoffmann, 2000 ). Indeed, some insects have the ability to detect contaminants in their food, leading them to adjust their consumption, which in turn results in lower mass (Nummelin et al., 2007 ; van Capelleveen et al., 1986 ; Vlahović et al., 2014 ). These findings are similar to Drosophila melanogaster that showed an aversion to cadmium-contaminated food (Bahadorani and Hilliker, 2009 ), although this adaptive strategy would be viable only on the short term. While we observed differences in food consumption between treated and control groups, these differences were unexpected as treated larvae consumed more than the controls at the approach of the pre-pupal stage. As observed in the control group, healthy individuals generally consume less when preparing for metamorphosis, often by purging just before forming the chrysalis (Kraus et al., 2014 ). However, in instances where development is delayed, as it has been shown with cadmium (Humann-Guilleminot et al., 2024 ), this reduction in food consumption may also occurs later than usual. Interestingly, the increased food consumption observed in the treated groups did not translate into mass gain, suggesting a physiological cost associated with the body's detoxification processes. Another possible explanation for this phenomenon is that ingested cadmium is predominantly stored in the insects' guts, which may impact their digestive enzymes, affecting nutrient absorption (Hensbergen et al., 2000 ; Su et al., 2023 ; Zhang et al., 2020 ). At the larval stage, we observed a trend toward cadmium accumulation, reaching a saturation threshold in the pupal stage, followed by a decrease in the adult stage. These results are consistent with observations in Lymantria dispar , where cadmium levels continuously increase during larval development, peaking at the pupal stage, and then decreasing in adults (Gintenreiter et al., 1993 ). This suggests that a significant amount of cadmium was eliminated during metamorphosis, particularly after exposure to higher levels. It has been shown under these conditions that metals are purged primarily during this phase, resulting in concentrations in adults up to 125 times lower than those in larvae (Kraus et al., 2014 ; Lindqvist and Block, 1995). Our results show that insects excrete a significant amount of cadmium through the integument and exuviae during metamorphosis, and do not absorb all ingested cadmium as evidenced by high concentrations in their feces. Numerous studies on insects from various orders and habitats have shown that cadmium accumulates mainly in their digestive tract at larval stage, serving as a major organ of protection against metals, by trapping them in epithelial cells (Aoki et al., 1984 ; Dallinger, 1993 ; Kafel et al., 2003 ). However, the cadmium absorbed by the midgut is then transported by the haemolymph and distributed in all parts of the body (Kafel et al., 2003 ; Leonard et al., 2009 ) which probably explains the concentrations measured here in the heads of larvae and adults, and in the antennae. In addition, cadmium stored in the meconium – a metabolic waste– has been shown to be excreted upon adult emergence (Aoki and Suzuki, 1984 ; Dallinger, 1993 ; Vlahović et al., 2017 ), which could explain the significant reduction in cadmium levels observed in S. littoralis adults compared to the pre-pupal stage (Dallinger, 1993 ; Dar et al., 2017 ; Lindqvist and Block, 1995). Altogether, these results highlight the critical role of cadmium excretion processes during the larval stages (feces, integument), metamorphosis (exuviae) and in adults in S. littoralis , and in insects in general. 5. Conclusion We have demonstrated the effects of nine cadmium concentrations on food consumption and larval growth in the noctuid moth Spodoptera littoralis . Our results show that cadmium concentrations peak during the pupal stage and then decrease in adults, indicating effective detoxification mechanisms during metamorphosis. This is confirmed by the significant amounts of cadmium excreted in feces, integument, and exuviae at the prepupal stage. In addition, we observe that larvae stored most of the ingested cadmium in their digestive tract, from where it was transferred by haemolymph to various body parts, including the head and the antennae. These results confirm the toxicity of cadmium and its role as an environmental stress factor. We recommend further research on the chronic effects of cadmium, with particular emphasis on the impact of contamination by sublethal doses throughout the food chain, at different trophic levels, considering potential synergistic impacts with other environmental pollutant. Declarations Ethical statement: We hereby confirm that this material is the authors' original work, has not been previously published, and accurately reflects their own research and analysis. The paper credits the significant contributions of co-authors and collaborators, and the results are properly contextualized within prior and existing research. All authors have been actively involved in the substantial work leading to this paper and take full public responsibility for its content. Funding: The project was supported by an Emergence funding program of Sorbonne University. Authors’ contributions Ségolène Humann-Guilleminot: Data curation, Formal analysis, Visualization, Writing – original draft, Writing – review & editing. Marie-Anne Pottier: Conceptualization, Investigation, Methodology. Maryse Rouelle: Investigation, Writing – review & editing. Annick Maria: Methodology, Resource. Martine Maibeche: Conceptualization, Funding acquisition, Project administration, Supervision, Writing – review & editing. David Siaussat: Conceptualization, Funding acquisition, Project administration, Supervision, Writing – review & editing. Data Availability All data used in this study is included within its Supplementary Information files. References Abedi, T., Mojiri, A., 2020. Cadmium uptake by wheat (Triticum aestivum L.): An overview. Plants 9. https://doi.org/10.3390/plants9040500 Ali, H., Khan, E., 2018. Bioaccumulation of non-essential hazardous heavy metals and metalloids in freshwater fish. Risk to human health. Environmental Chemistry Letters 16, 903–917. https://doi.org/10.1007/s10311-018-0734-7 Ali, H., Khan, E., Ilahi, I., 2019. Environmental chemistry and ecotoxicology of hazardous heavy metals: environmental persistence, toxicity, and bioaccumulation. Journal of Chemistry 2019, 1–301. https://doi.org/10.1155/2019/6730305 Aoki, Y., Suzuki, K.T., 1984. Excretion of cadmium and change in the relative ratio of iso-cadmium-binding proteins during metamorphosis of fleshfly (Sarcophaga peregrina). Comparative Biochemistry and Physiology Part C: Comparative Pharmacology 78, 315–317. https://doi.org/10.1016/0742-8413(84)90089-6 Aoki, Y., Suzuki, K.T., Kubota, K., 1984. Accumulation of cadmium and induction of its binding protein in the digestive tract of fleshfly (Sarcophaga peregrina) larvae. Comparative Biochemistry and Physiology Part C: Comparative Pharmacology 77, 279–282. https://doi.org/10.1016/0742-8413(84)90013-6 Aviles, A., Boulogne, I., Durand, N., Maria, A., Cordeiro, A., Bozzolan, F., Goutte, A., Alliot, F., Dacher, M., Renault, D., Maibeche, M., Siaussat, D., 2019. Effects of DEHP on post-embryonic development, nuclear receptor expression, metabolite and ecdysteroid concentrations of the moth Spodoptera littoralis. Chemosphere 215, 725–738. https://doi.org/10.1016/j.chemosphere.2018.10.102 Bahadorani, S., Hilliker, A.J., 2009. Biological and behavioral effects of heavy metals in Drosophila melanogaster adults and larvae. Journal of Insect Behavior 22, 399–411. https://doi.org/10.1007/s10905-009-9181-4 Bradl, H.B., 2005. Chapter 1 Sources and origins of heavy metals, in: Bradl, H.B. (Ed.), Interface Science and Technology. Elsevier, pp. 1–27. https://doi.org/10.1016/S1573-4285(05)80020-1 Brown, E.S., Dewhurst, C.F., 1975. The genus Spodoptera (Lepidoptera, Noctuidae) in Africa and the near East. Bulletin of Entomological Research 65, 221–262. https://doi.org/10.1017/S0007485300005939 Brunn, H., Arnold, G., Körner, W., Rippen, G., Steinhäuser, K.G., Valentin, I., 2023. PFAS: forever chemicals—persistent, bioaccumulative and mobile. Reviewing the status and the need for their phase out and remediation of contaminated sites. Environmental Sciences Europe 35, 20. https://doi.org/10.1186/s12302-023-00721-8 Butt, A., Qurat ul, A., Rehman, K., Khan, M.X., Hesselberg, T., 2018. Bioaccumulation of cadmium, lead, and zinc in agriculture-based insect food chains. Environmental Monitoring and Assessment 190, 698. https://doi.org/10.1007/s10661-018-7051-2 Byers, J.R., Hinks, C.F., 1976. Biosystematics of the genus Euxoa (Lepidoptera: Noctuidae): V. Rearing Procedures, and life cycles of 36 species. The Canadian Entomologist 108, 1345–1357. https://doi.org/10.4039/Ent1081345-12 Castañeda, L.E., Figueroa, C.C., Fuentes-Contreras, E., Niemeyer, H.M., Nespolo, R.F., 2009. Energetic costs of detoxification systems in herbivores feeding on chemically defended host plants: a correlational study in the grain aphid, Sitobion avenae. The Journal of experimental biology 212, 1185–90. https://doi.org/10.1242/jeb.020990 Cervera, A., Maymó, A.C., Sendra, M., Martínez-Pardo, R., Garcerá, M.D., 2004. Cadmium effects on development and reproduction of Oncopeltus fasciatus (Heteroptera: Lygaeidae). Journal of Insect Physiology 50, 737–749. https://doi.org/10.1016/j.jinsphys.2004.06.001 Conolly, R.B., Lutz, W.K., 2004. Nonmonotonic dose-response relationships: mechanistic basis, kinetic modeling, and implications for risk assessment. Toxicological Sciences 77, 151–157. https://doi.org/10.1093/toxsci/kfh007 Dallinger, R., 1993. Strategies of metal detoxification in terestrial invertebrates. Ecotoxicology of Metals in Invertebrates 245–290. Dar, M.I., Green, I.D., Naikoo, M.I., Khan, F.A., Ansari, A.A., Lone, M.I., 2017. Assessment of biotransfer and bioaccumulation of cadmium, lead and zinc from fly ash amended soil in mustard–aphid–beetle food chain. Science of The Total Environment 584–585, 1221–1229. https://doi.org/10.1016/j.scitotenv.2017.01.186 Farrar, R.R., Barbour, J.D., Kennedy, G.G., 1989. Quantifying food consumption and growth in insects. Annals of the Entomological Society of America 82, 593–598. https://doi.org/10.1093/aesa/82.5.593 Frat, L., Chertemps, T., Pesce, E., Bozzolan, F., Dacher, M., Planelló, R., Herrero, O., Llorente, L., Moers, D., Siaussat, D., 2021. Single and mixed exposure to cadmium and mercury in Drosophila melanogaster: Molecular responses and impact on post-embryonic development. Ecotoxicology and Environmental Safety 220, 112377. https://doi.org/10.1016/j.ecoenv.2021.112377 Gao, D.-W., Wen, Z.-D., 2016. Phthalate esters in the environment: A critical review of their occurrence, biodegradation, and removal during wastewater treatment processes. Science of The Total Environment 541, 986–1001. https://doi.org/10.1016/j.scitotenv.2015.09.148 Gintenreiter, S., Ortel, J., Nopp, H.J., 1993. Effects of different dietary levels of cadmium, lead, copper, and zinc on the vitality of the forest pest insect Lymantria dispar L. (Lymantriidae, Lepid). Archives of Environmental Contamination and Toxicology 25, 62–66. https://doi.org/10.1007/BF00230712 Haider, F.U., Liqun, C., Coulter, J.A., Cheema, S.A., Wu, J., Zhang, R., Wenjun, M., Farooq, M., 2021. Cadmium toxicity in plants: Impacts and remediation strategies. Ecotoxicology and Environmental Safety 211, 111887. https://doi.org/10.1016/j.ecoenv.2020.111887 Health, E.P. on P., 2015. Scientific Opinion on the pest categorisation of Spodoptera littoralis. EFSA Journal 13, 3987. https://doi.org/10.2903/j.efsa.2015.3987 Hensbergen, P.J., van Velzen, M.J.M., Adi Nugroho, R., Donker, M.H., van Straalen, N.M., 2000. Metallothionein-bound cadmium in the gut of the insect Orchesella cincta (Collembola) in relation to dietary cadmium exposure. Comparative Biochemistry and Physiology Part C: Pharmacology, Toxicology and Endocrinology 125, 17–24. https://doi.org/10.1016/S0742-8413(99)00087-0 Humann-Guilleminot, S., Fuentes, A., Maria, A., Couzi, P., Siaussat, D., 2024. Cadmium and phthalate impact developmental growth and mortality of Spodoptera littoralis, but not reproductive success. Ecotoxicology and Environmental Safety 281, 116605. https://doi.org/10.1016/j.ecoenv.2024.116605 Iavicoli, I., Fontana, L., Bergamaschi, A., 2009. The Effects of metals as endocrine disruptors. Journal of Toxicology and Environmental Health, Part B 12, 206–223. https://doi.org/10.1080/10937400902902062 Jin, R., Bu, D., Liu, G., Zheng, M., Lammel, G., Fu, J., Yang, L., Li, C., Habib, A., Yang, Y., Liu, X., 2020. New classes of organic pollutants in the remote continental environment – Chlorinated and brominated polycyclic aromatic hydrocarbons on the Tibetan Plateau. Environment International 137, 105574. https://doi.org/10.1016/j.envint.2020.105574 Kafel, A., Bednarska, K., Augustyniak, M., Witas, I., Szuliñska, E., 2003. Activity of glutathione S-transferase in Spodoptera exigua larvae exposed to cadmium and zinc in two subsequent generations. Environment International 28, 683–686. https://doi.org/10.1016/S0160-4120(02)00111-3 Kraus, J.M., Walters, D.M., Wesner, J.S., Stricker, C.A., Schmidt, T.S., Zuellig, R.E., 2014. Metamorphosis Alters Contaminants and Chemical Tracers in Insects: Implications for Food Webs. Environ. Sci. Technol. 48, 10957–10965. https://doi.org/10.1021/es502970b Kubier, A., Wilkin, R.T., Pichler, T., 2019. Cadmium in soils and groundwater: A review. Applied Geochemistry 108, 104388. https://doi.org/10.1016/j.apgeochem.2019.104388 Kuznetsova, A., Brockhoff, P.B., Christensen, R.H.B., 2017. lmerTest package: tests in linear mixed effects models. Journal of Statistical Software 82, 26. https://doi.org/10.18637/jss.v082.i13 Leonard, E.M., Pierce, L.M., Gillis, P.L., Wood, C.M., O’Donnell, M.J., 2009. Cadmium transport by the gut and Malpighian tubules of Chironomus riparius. Aquatic Toxicology 92, 179–186. https://doi.org/10.1016/j.aquatox.2009.01.011 Lindqvist, L., Block, M., 1995. Excretion of cadmium during moulting and metamorphosis in Tenebrio molitor (Coleoptera; Tenebrionidae). Comparative Biochemistry and Physiology Part C: Pharmacology, Toxicology and Endocrinology 111, 325–328. https://doi.org/10.1016/0742-8413(95)00057-U Lüdecke, D., Ben Shachar, M., Patil, I., Waggoner, P., Makowski, D., 2021. Performance: an R package for assessment, comparison and testing of statistical models. The Journal of Open Source Software 6, 3139. https://doi.org/10.21105/joss.03139 Marcus, S.R., Fiumera, A.C., 2016. Atrazine exposure affects longevity, development time and body size in Drosophila melanogaster. Journal of Insect Physiology 91–92, 18–25. https://doi.org/10.1016/j.jinsphys.2016.06.006 Maryanski, M., Kramarz, P., Laskowski, R., Niklinska, M., 2002. Decreased Energetic Reserves, Morphological Changes and Accumulation of Metals in Carabid Beetles (Poecilus cupreus L.) Exposed to Zinc- or Cadmium-contaminated Food. Ecotoxicology 11, 127–139. https://doi.org/10.1023/A:1014425113481 Migula, P., Binkowska, K., 1993. Feeding strategies of grasshoppers (Chorthippus sp.) on heavy metal contaminated plants. Science of The Total Environment 134, 1071–1083. https://doi.org/10.1016/S0048-9697(05)80112-3 Moulis, J.-M., 2010. Cellular mechanisms of cadmium toxicity related to the homeostasis of essential metals. BioMetals 23, 877–896. https://doi.org/10.1007/s10534-010-9336-y Nagorka, R., Koschorreck, J., 2020. Trends for plasticizers in German freshwater environments – Evidence for the substitution of DEHP with emerging phthalate and non-phthalate alternatives. Environmental Pollution 262, 114237. https://doi.org/10.1016/j.envpol.2020.114237 Net, S., Sempéré, R., Delmont, A., Paluselli, A., Ouddane, B., 2015. Occurrence, fate, behavior and ecotoxicological state of phthalates in different environmental matrices. Environmental Science & Technology 49, 4019–4035. https://doi.org/10.1021/es505233b Nummelin, M., Lodenius, M., Tulisalo, E., Hirvonen, H., Alanko, T., 2007. Predatory insects as bioindicators of heavy metal pollution. Environmental Pollution 145, 339–347. https://doi.org/10.1016/j.envpol.2006.03.002 Ortel, J., 1995. Effects of metals on the total lipid content in the gypsy moth (Lymantria dispar, lymantriidae, lepid.) and its hemolymph. Bulletin of Environmental Contamination and Toxicology 55, 216–221. https://doi.org/10.1007/BF00203012 Planelló, R., Martínez-Guitarte, J.L., Morcillo, G., 2010. Effect of acute exposure to cadmium on the expression of heat-shock and hormone-nuclear receptor genes in the aquatic midge Chironomus riparius. Science of The Total Environment 408, 1598–1603. https://doi.org/10.1016/j.scitotenv.2010.01.004 Quimby, P.C., Frick, K.E., Wauchope, R.D., Kay, S.H., 1979. Effects of cadmium on two biocontrol insects and their host weeds. Bulletin of Environmental Contamination and Toxicology 22, 371–378. https://doi.org/10.1007/BF02026957 R Core Team, 2022. R: A language and environment for statistical computing. [WWW Document]. Raina, R.M., Pawar, P., Sharma, R.N., 2001. Developmental inhibition and reproductive potential impairment in Musca domestica L. by heavy metals. Indian journal of experimental biology 39, 78–81. Raise, G., Gemmellaro, M.D., 2024. A review on the effects of heavy metals on the development of blow flies (Diptera: Calliphoridae). WIREs Forensic Science 6, e1503. https://doi.org/10.1002/wfs2.1503 Rana, S.V.S., 2014. Perspectives in endocrine toxicity of heavy metals—A Review. Biological Trace Element Research 160, 1–14. https://doi.org/10.1007/s12011-014-0023-7 Rathi, B.S., Kumar, P.S., Vo, D.-V.N., 2021. Critical review on hazardous pollutants in water environment: Occurrence, monitoring, fate, removal technologies and risk assessment. Science of The Total Environment 797, 149134. https://doi.org/10.1016/j.scitotenv.2021.149134 Schmidt, G.H., Ibrahim, N.M.M., Abdallah, M.D., 1992. Long-term effects of heavy metals in food on developmental stages of Aiolopus thalassinus (Saltatoria: Acrididae). Archives of Environmental Contamination and Toxicology 23, 375–382. https://doi.org/10.1007/BF00216248 Sildanchandra, W., Crane, M., 2000. Influence of sexual dimorphism in Chironomus riparius Meigen on toxic effects of cadmium. Environmental Toxicology and Chemistry 19, 2309–2313. https://doi.org/10.1002/etc.5620190921 Sorvari, J., Rantala, L.M., Rantala, M.J., Hakkarainen, H., Eeva, T., 2007. Heavy metal pollution disturbs immune response in wild ant populations. Environmental Pollution 145, 324–328. https://doi.org/10.1016/j.envpol.2006.03.004 Su, H., Wu, M., Yang, Yong, Deng, Y., Yang, Yi-zhong, Sun, Q., 2023. Tissue distribution of cadmium and its effect on reproduction in Spodoptera exigua. Journal of Integrative Agriculture. https://doi.org/10.1016/j.jia.2023.08.010 Tabassum, S., Kotnala, C.B., Salman, M., Tariq, M., Khan, A.H., Khan, N.A., 2024. The impact of heavy metal concentrations on aquatic insect populations in the Asan Wetland of Dehradun, Uttarakhand. Scientific Reports 14, 4824. https://doi.org/10.1038/s41598-024-52522-5 van Capelleveen, H.E., van Straalen, N.M., van den Berg, M., van Wachem, E. (Eds.), 1986. Avoidance as a mechanism of tolerance for lead in terrestrial arthropods. Nederlandse Entomologische Vereniging, Amsterdam, The Netherlands. van Straalen, N.M., Hoffmann, A.A., 2000. Review of experimental evidence for physiological costs of tolerance to toxicants., in: Demography in Ecotoxicology. John Wiley and Sons, pp. 147–161. Vandenberg, L.N., Colborn, T., Hayes, T.B., Heindel, J.J., Jacobs, D.R., Jr., Lee, D.H., Shioda, T., Soto, A.M., vom Saal, F.S., Welshons, W.V., Zoeller, R.T., Myers, J.P., 2012. Hormones and endocrine-disrupting chemicals: low-dose effects and nonmonotonic dose responses. Endocrine reviews 33, 378–455. https://doi.org/10.1210/er.2011-1050 Vardhan, K.H., Kumar, P.S., Panda, R.C., 2019. A review on heavy metal pollution, toxicity and remedial measures: Current trends and future perspectives. Journal of Molecular Liquids 290, 111197. https://doi.org/10.1016/j.molliq.2019.111197 Verma, N., Rachamalla, M., Kumar, P.S., Dua, K., 2023. Chapter 6 - Assessment and impact of metal toxicity on wildlife and human health, in: Shukla, S.K., Kumar, S., Madhav, S., Mishra, P.K. (Eds.), Metals in Water. Elsevier, pp. 93–110. https://doi.org/10.1016/B978-0-323-95919-3.00002-1 Vlahović, M., Ilijin, L., Lazarević, J., Mrdaković, M., Gavrilović, A., Matić, D., Mataruga, V.P., 2014. Cadmium-induced changes of gypsy moth larval mass and protease activity. Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology 160, 9–14. https://doi.org/10.1016/j.cbpc.2013.11.002 Vlahović, M., Matić, D., Mutić, J., Trifković, J., Đurđić, S., Perić Mataruga, V., 2017. Influence of dietary cadmium exposure on fitness traits and its accumulation (with an overview on trace elements) in Lymantria dispar larvae. Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology 200, 27–33. https://doi.org/10.1016/j.cbpc.2017.06.003 Wasserstein, R.L., Lazar, N.A., 2016. The ASA Statement on p-Values: Context, Process, and Purpose. The American Statistician 70, 129–133. https://doi.org/10.1080/00031305.2016.1154108 Wu, G.-X., Ye, G.-Y., Hu, C., Cheng, J.-A., 2006. Accumulation of cadmium and its effects on growth, development and hemolymph biochemical compositions in Boettcherisca peregrina larvae (Diptera: Sarcophagidae). Insect Science 13, 31–39. https://doi.org/10.1111/j.1744-7917.2006.00065.x Zhang, Y., Wolosker, M.B., Zhao, Y., Ren, H., Lemos, B., 2020. Exposure to microplastics cause gut damage, locomotor dysfunction, epigenetic silencing, and aggravate cadmium (Cd) toxicity in Drosophila. Science of The Total Environment 744, 140979. https://doi.org/10.1016/j.scitotenv.2020.140979 Zulfiqar, U., Jiang, W., Xiukang, W., Hussain, S., Ahmad, M., Maqsood, M.F., Ali, N., Ishfaq, M., Kaleem, M., Haider, F.U., Farooq, N., Naveed, M., Kucerik, J., Brtnicky, M., Mustafa, A., 2022. Cadmium phytotoxicity, tolerance, and advanced remediation approaches in agricultural soils; A comprehensive review. Frontiers in Plant Science 13. https://doi.org/10.3389/fpls.2022.773815 Additional Declarations No competing interests reported. Supplementary Files CadmiumFoodConsoDS.xlsx CadmiumLarvalMassDS.xlsx CadmiumdosageBodyPart.xlsx CadmiumdosagedevelopmentDS.xlsx LarvalmortalityDS.xlsx SuppMatCadmiumdosagegrowthSpodopteraLittoralisV1SHG.docx Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-5439205","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":381245607,"identity":"16468132-cbcf-477c-bfe2-4fb9b3e60d96","order_by":0,"name":"Ségolène Humann-Guilleminot","email":"","orcid":"","institution":"Sorbonne Université, Univ Paris Est Créteil, CNRS, INRAE, IRD","correspondingAuthor":false,"prefix":"","firstName":"Ségolène","middleName":"","lastName":"Humann-Guilleminot","suffix":""},{"id":381245608,"identity":"fa1f54e8-5849-49c2-8b5d-e6d1207d5733","order_by":1,"name":"Marie-Anne Pottier","email":"","orcid":"","institution":"Sorbonne Université, Univ Paris Est Créteil, CNRS, INRAE, IRD","correspondingAuthor":false,"prefix":"","firstName":"Marie-Anne","middleName":"","lastName":"Pottier","suffix":""},{"id":381245609,"identity":"6bc46c0f-e0f3-4fb8-9b4f-4cb23b12cca3","order_by":2,"name":"Maryse Rouelle","email":"","orcid":"","institution":"Milieux Environnementaux, Transferts et Interactions dans les hydrosystèmes et les Sols METIS – CNRS – EPHE, UMR 7619","correspondingAuthor":false,"prefix":"","firstName":"Maryse","middleName":"","lastName":"Rouelle","suffix":""},{"id":381245610,"identity":"f3ae251b-0651-48cc-be17-928ae0481f8c","order_by":3,"name":"Annick Maria","email":"","orcid":"","institution":"Sorbonne Université, Univ Paris Est Créteil, CNRS, INRAE, IRD","correspondingAuthor":false,"prefix":"","firstName":"Annick","middleName":"","lastName":"Maria","suffix":""},{"id":381245611,"identity":"02f01831-135a-4196-b519-697b4702e40d","order_by":4,"name":"Martine Maïbèche","email":"","orcid":"","institution":"Sorbonne Université, Univ Paris Est Créteil, CNRS, INRAE, IRD","correspondingAuthor":false,"prefix":"","firstName":"Martine","middleName":"","lastName":"Maïbèche","suffix":""},{"id":381245612,"identity":"60051386-2c7b-4d8e-addb-0042b22eac54","order_by":5,"name":"David Siaussat","email":"data:image/png;base64,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","orcid":"","institution":"Sorbonne Université, Univ Paris Est Créteil, CNRS, INRAE, IRD","correspondingAuthor":true,"prefix":"","firstName":"David","middleName":"","lastName":"Siaussat","suffix":""}],"badges":[],"createdAt":"2024-11-12 11:38:25","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-5439205/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-5439205/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":70363187,"identity":"d3627238-0660-4b24-a253-3f9397b8d999","added_by":"auto","created_at":"2024-12-02 13:49:27","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":1352847,"visible":true,"origin":"","legend":"\u003cp\u003eFood consumption (%) and mass of individuals over six days, segmented by treatment, along with the relative growth rate for each treatment and the percentage of larval mortality. Grey dots represent the mass, food consumption, or relative growth rate (RGR) of each individual on each day for each treatment. Coloured dots indicate the averages of these variables, and the corresponding lines connect the mean dots, illustrating the variations among the different treatments. Note: the barplot is for illustrative purposes only and does not strictly reflect the statistical analysis conducted.\u003c/p\u003e","description":"","filename":"Figure120.png","url":"https://assets-eu.researchsquare.com/files/rs-5439205/v1/f658f78872dc5dd91edeb976.png"},{"id":70362912,"identity":"5fca79cd-1d3f-452f-977f-ed911aa6ffc4","added_by":"auto","created_at":"2024-12-02 13:41:27","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":313400,"visible":true,"origin":"","legend":"\u003cp\u003eCadmium concentration at different stages of development in \u003cem\u003eSpodoptera littoralis\u003c/em\u003e. The graph shows cadmium concentrations (µg/g) in fourth, fifth and sixth larval stages (L4 to L6), pupae, and adults from three treatment groups: Control (0 µg/g), low cadmium (15.625 µg/g), and high cadmium (125 µg/g) treatment. Mean cadmium concentrations are depicted for each stage, with jittered individual grey data points overlaid. All concentrations are in µg/g of dry weight.\u003c/p\u003e","description":"","filename":"Figure217.png","url":"https://assets-eu.researchsquare.com/files/rs-5439205/v1/c728b0ac19655f99766343e7.png"},{"id":70364740,"identity":"afa4c427-b9fe-4e76-bbc8-fdd35a67488e","added_by":"auto","created_at":"2024-12-02 13:57:27","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":391524,"visible":true,"origin":"","legend":"\u003cp\u003eCadmium concentration in different body parts in larval, pre-pupal and adult stages of \u003cem\u003eSpodoptera littoralis\u003c/em\u003e. The graph shows cadmium concentrations (µg/g) in larval, pupal, and adults from three treatment groups: Control (0 µg/g), medium cadmium (62.5 µg/g), and high cadmium (125 µg/g). Mean cadmium concentrations are depicted for each stage, with individual grey data points overlaid. All concentrations are in µg/g of dry weight except for haemolymph that is in µg/g of fresh weight.\u003c/p\u003e","description":"","filename":"Figure311.png","url":"https://assets-eu.researchsquare.com/files/rs-5439205/v1/f18135652e5ed4b47ecd46db.png"},{"id":78766279,"identity":"28df1b87-bd0e-4dca-abdc-ce96ac7aa5b4","added_by":"auto","created_at":"2025-03-18 15:02:06","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":3712105,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5439205/v1/d8f79d82-b079-4270-a2df-da1e7c6f662d.pdf"},{"id":70362917,"identity":"c91d225e-62d4-400a-b248-3dae6798b022","added_by":"auto","created_at":"2024-12-02 13:41:27","extension":"xlsx","order_by":5,"title":"","display":"","copyAsset":false,"role":"supplement","size":40894,"visible":true,"origin":"","legend":"","description":"","filename":"CadmiumFoodConsoDS.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-5439205/v1/9531f594990e609e0c36a4f9.xlsx"},{"id":70363189,"identity":"5ce03582-42a0-43b6-bbf2-402c131f6e4a","added_by":"auto","created_at":"2024-12-02 13:49:27","extension":"xlsx","order_by":6,"title":"","display":"","copyAsset":false,"role":"supplement","size":33920,"visible":true,"origin":"","legend":"","description":"","filename":"CadmiumLarvalMassDS.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-5439205/v1/c3c26a71e162eb582ef7d9ba.xlsx"},{"id":70365152,"identity":"c8a4f9d0-5726-4cee-9b92-036570e212fe","added_by":"auto","created_at":"2024-12-02 14:05:27","extension":"xlsx","order_by":7,"title":"","display":"","copyAsset":false,"role":"supplement","size":14709,"visible":true,"origin":"","legend":"","description":"","filename":"CadmiumdosageBodyPart.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-5439205/v1/00ebbe1985ac7748a589fff4.xlsx"},{"id":70364739,"identity":"6246b405-485c-4651-b468-bde940af9d99","added_by":"auto","created_at":"2024-12-02 13:57:27","extension":"xlsx","order_by":8,"title":"","display":"","copyAsset":false,"role":"supplement","size":16583,"visible":true,"origin":"","legend":"","description":"","filename":"CadmiumdosagedevelopmentDS.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-5439205/v1/76cbc700886ae7e6bf850603.xlsx"},{"id":70362915,"identity":"235d425d-961d-4bf2-8187-80d6726e8f7a","added_by":"auto","created_at":"2024-12-02 13:41:27","extension":"xlsx","order_by":9,"title":"","display":"","copyAsset":false,"role":"supplement","size":12275,"visible":true,"origin":"","legend":"","description":"","filename":"LarvalmortalityDS.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-5439205/v1/91cec51d0d08985b28ad3c20.xlsx"},{"id":70363192,"identity":"3913f196-9870-4068-8c34-364ae263f68b","added_by":"auto","created_at":"2024-12-02 13:49:27","extension":"docx","order_by":10,"title":"","display":"","copyAsset":false,"role":"supplement","size":46672,"visible":true,"origin":"","legend":"","description":"","filename":"SuppMatCadmiumdosagegrowthSpodopteraLittoralisV1SHG.docx","url":"https://assets-eu.researchsquare.com/files/rs-5439205/v1/c25b8139929fa15b42fa6696.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Dose-dependent effects of cadmium on food consumption, mass, and bioaccumulation in the moth Spodoptera littoralis","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eIn recent decades, the accumulation of environmental pollutants in various ecosystems, largely due to industrial and agricultural activities, has raised growing concern (Gao and Wen, \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Jin et al., \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Nagorka and Koschorreck, \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Vardhan et al., \u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Among pollutants, heavy metals accumulate at varying concentrations in the environment and in animals, resulting in a wide range of physiological effects (Ali and Khan, \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Tabassum et al., \u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Verma et al., \u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Due to their toxicity, persistence and bioaccumulation capacity, they pose significant long-term risks as they can remain in ecosystems for centuries (Ali et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Brunn et al., \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Net et al., \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2015\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eCadmium, a heavy metal element, is released into the environment from various sources including industrial wastes and agricultural runoff (Bradl, \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2005\u003c/span\u003e, Kubier et al., \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Like other metals, cadmium is persistent in the environment and accumulates in living organisms, with potential adverse effects on growth and acting as an endocrine disruptor (Ali and Khan, \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Raina et al., \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e2001\u003c/span\u003e; Sorvari et al., \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e2007\u003c/span\u003e; Verma et al., \u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). The toxic effects of cadmium are well-documented and affect various physiological processes in microorganisms, plants and animals. However, its impact varies by species and developmental stages, with some organisms showing a higher propensity to bioaccumulate and adverse effects (Vardhan et al., \u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). For instance, cadmium is known to alter the expression of hormone receptor genes and to disrupt circulating hormone levels, which has negative effects on moulting, growth and reproduction in various invertebrate species (Iavicoli et al., \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2009\u003c/span\u003e; Planell\u0026oacute; et al., \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Rana, \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). In several insect species, such as the flies \u003cem\u003eBoettcherisca peregrina, Drosophila melanogaster\u003c/em\u003e and \u003cem\u003eMusca domestica\u003c/em\u003e, high doses of cadmium ingestion delay the transitions from larval to adult development, and similar findings were reported for mercury (Frat et al., \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Raina et al., \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e2001\u003c/span\u003e; Wu et al., \u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e2006\u003c/span\u003e). In addition, studies have shown a detrimental negative impact of cadmium and other heavy metals on mass and survival of blow flies, particularly in larval stages (review in Raise and Gemmellaro, \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2024\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIn a previous study on the model pest moth \u003cem\u003eSpodoptera littoralis\u003c/em\u003e, we showed that ingestion of cadmium at low and high (sublethal) doses delayed post-embryonic development and affected larval growth and survival (Humann-Guilleminot et al., \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). As a widespread agricultural pest, \u003cem\u003eS. littoralis\u003c/em\u003e interacts closely with various ecosystems and is frequently exposed to different pollutants (EFSA, 2015). Its short life cycle and well-documented developmental stages make it an ideal model for studying the effects of pollutants over a full life cycle (Brown and Dewhurst, \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e1975\u003c/span\u003e). In addition, its physiological and developmental processes are similar to those of other insects, allowing the findings to be extrapolated into a broader ecological context. To better understand the toxicity of cadmium in this species, we experimentally evaluated the effects on dietary intake of nine doses of cadmium, ranging from sublethal to lethal, administered orally to larvae, on their food consumption, mass gain, relative growth rate, and mortality over a 6-day period. We also examined the bioaccumulation of two sublethal doses of ingested cadmium at different developmental stages, from the fourth larval stage to adulthood. We studied the distribution of cadmium in the tissues of final larval and pre-pupal stages, and measured its concentrations in tissues and haemolymph of adults. We hypothesized that cadmium would have a negative impact on all measured variables, including food intake, mass gain, relative growth rate and larval mortality, with more pronounced effects at higher doses. In addition, we expected that cadmium would be detected in all individuals that ingested it, with higher concentrations corresponding to higher doses. We also predicted an increase in the concentration of cadmium at the larval stage, followed by a decrease at adulthood, due to detoxification during metamorphosis.\u003c/p\u003e"},{"header":"2. Material and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1. Moths rearing and exposure to treatment\u003c/h2\u003e \u003cp\u003e \u003cem\u003eS. littoralis\u003c/em\u003e larvae were reared on a semi-artificial diet, as described in (Humann-Guilleminot et al., \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). They were stored in glass jars at 4\u0026deg;C until use, in order to ensured minimal chemical contamination, as both treated and control diets were stored in glass rather than plastic containers. Larvae were raised in DEHP-free plastic boxes, which were replaced regularly for each experiment. All laboratory materials have been sterilized to avoid microbial contamination and excessive mortality. The uniform distribution and absorption of cadmium in the diet was achieved by incorporating cadmium (CdCl\u003csub\u003e2٠\u003c/sub\u003e 2.5H\u003csub\u003e2\u003c/sub\u003eO, 239208-100G, Sigma, France) into food that was in liquid form. The mixture has been mixed with an industrial stirrer for an extended period. Agar was included to solidify the mixture into a uniform jelly after cooling and then consumed by insect larvae. From the fourth larval instar to pupation (corresponding to about 15 days), larvae were fed with food containing nine cadmium concentrations (\u0026ldquo;T1\u0026rdquo; to \u0026ldquo;T9\u0026rdquo;) prepared from a 3M solution, i.e. 7.8, 15.625, 31.25, 62.5, 125, 250, 500 \u0026micro;g, 5 and 50 mg CdCl\u003csub\u003e2\u003c/sub\u003e per gram of food. Control larvae received a diet mixed with solvent solution (500 \u0026micro;L of ethanol in 100 g semi-solid food). Larvae were fed individually ad-libitum under specific contamination conditions.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2. Assessment of larval food consumption, growth and mortality\u003c/h2\u003e \u003cp\u003eThe experimental phase included 200 larvae (N=20 per condition). The food was weighed daily to calculate the rate of consumption according to the CdCl\u003csub\u003e2\u003c/sub\u003e concentration, and the larvae were also weighed daily until pupation. The relative growth rate was calculated using the formula from Farrar et al., 1989: \u0026nbsp;\u003cimg src=\"https://myfiles.space/user_files/127393_c7e80a1c9bb65875/127393_custom_files/img1733146641.png\"\u003e\u0026nbsp;(average mass at day 1 and day 6). Finally, the mortality rates at the larval and pupal stages were recorded for each treatment.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.3. Cadmium quantification\u003c/h2\u003e \u003cp\u003eTo evaluate cadmium bioaccumulation, we quantified cadmium in treated and control insects using an atomic absorption spectrometer. Samples were analysed after a mineralization process in a Teflon flask at 100\u0026deg;C. They were mixed with 1 ml of 68% nitric acid (HNO₃; VWR Analar Normapur) for 24 hours, then with 0.5 ml of 30% hydrogen peroxide (H₂O₂; VWR Analar Normapur) for again 24 hours. Acid mineralization was performed to break down the sample matrix, releasing the elements into a solution for further analysis. Once mineralized, the liquid samples were transferred into tubes (215\u0026ndash;0329 from VWR) and diluted with 6 ml of MilliQ water (Purelab Ultra system from Elga). The weight of each tube was recorded for precise measurement of its content. Depending on the analyses, the insects were fed different doses of cadmium and different tissues, or body parts were dissected to perform the measurements. To follow cadmium concentrations over postembryonic development, two groups of larvae received either 15.625 (T2) or 125 \u0026micro;g (T5) of CdCl\u003csub\u003e2\u003c/sub\u003e from the 4th larval stage to adulthood. At least three insects were sampled daily for whole-body analysis. Larvae were fasted for 24 hours to clean their digestive tracts before any measurements. To evaluate cadmium concentration in different tissues at various stages of development, including final larval, pre-pupal and adult stages, two other larval groups were fed either 62.5 (T4) or 125 \u0026micro;g (T5) cadmium using set of individuals that differed from those used for whole-body measurements. For the final larval stage, we analysed the digestive tract, the fat body and the head. For the pre-pupal stage, cadmium levels were measured in the feces, in exuviae and integument. In male and female adults, measurements were made from the head, thorax, abdomen, antennae and haemolymph. All tissues were thoroughly rinsed in Ringer\u0026rsquo;s solution, especially the digestive tract. Haemolymph was collected from one-day-old adults, while larval feces were collected throughout the study. For each stage and each treatment, we analysed five samples in each category. To ensure sufficient analysis, tissues were pooled in triplicate to achieve a minimum of 50 mg of material, then dried at 150\u0026deg;C for 10 to 15 minutes. Exceptions to this drying protocol were haemolymph, antennae, and exuviae, which were handled differently. We then measured the quantity of cadmium in each pool using Inductively Coupled Plasma Optical Emission Spectrometry (ICP-OES Agilent 5100SDVD, Agilent Technologies, Santa Clara, CA, U.S.A.) at the ALIPP6 platform (Sorbonne University, Paris, France), with a detection threshold of ICP-OES at 3 mg/kg.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003e2.4. Statistical analyses\u003c/h2\u003e \u003cp\u003eAll statistical analyses were performed using the R v. 4.2.2 software (R Core Team, \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). For each analysis, the control group was set as the reference using the \u003cem\u003erelevel\u003c/em\u003e function from the \u003cem\u003estats\u003c/em\u003e package.\u003c/p\u003e \u003cdiv id=\"Sec7\" class=\"Section3\"\u003e \u003ch2\u003e2.4.1. Effect of cadmium on larval food consumption, mass, relative growth rate and mortality\u003c/h2\u003e \u003cp\u003eWe first aimed to assess the evolution of both larval food consumption and mass from the 4th larval stage to pre-pupae under ten different treatments (9 doses of cadmium and control). We ran two linear mixed effect models using the \u003cem\u003elmer\u003c/em\u003e function from the R package \u003cem\u003elmerTest\u003c/em\u003e (Kuznetsova et al., \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2017\u003c/span\u003e), that included either the logit-transformed percentage of food consumption or log-transformed body mass as the response variable, day (from 0 to 6, continuous), treatment (categorical), interaction between day and treatment, and quadratic term of day and interaction between quadratic term day and treatment to account for the observed non-linear relationship between time and food consumption, or mass, at larval stage day 0 relative to larval stage at day 6 as explanatory variables. The model included individual identity as random intercept to account for the repeated measures taken within each individual. We then conducted a linear model using the \u003cem\u003elm\u003c/em\u003e from the \u003cem\u003estats\u003c/em\u003e package to assess the influence of cadmium treatments on larvae relative growth rates, that included the relative growth rate as the response variable and the treatment as the explanatory variable. Lastly, we conducted a generalized linear model with a binomial distribution to assess the effect of treatments on larval mortality. The model was implemented using the \u003cem\u003eglm\u003c/em\u003e function from the \u003cem\u003estats\u003c/em\u003e package. The response variable was the binary outcome of larval mortality, with treatment as the explanatory variable. However, since larval mortality was 100% in the treatments with the three highest cadmium doses (T7, T8, T9), these treatments were excluded from the analysis.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section3\"\u003e \u003ch2\u003e2.4.2. Bioaccumulation of cadmium across developmental stages and tissues\u003c/h2\u003e \u003cp\u003eWe analysed the differences in bioaccumulation of two sublethal doses (T2 and T5) of ingested cadmium at different developmental stages and in various tissues. We first applied a linear model on a first set of data using the \u003cem\u003elm\u003c/em\u003e function from the \u003cem\u003estats\u003c/em\u003e package, with log-transformed cadmium concentration as the response variable, and stage (from the 4th to the 6th, pre-pupae, 3-day pupae, 9-day pupae, and adults), treatment, and the interaction between stage and treatment as explanatory variables. Furthermore, we investigated cadmium concentrations in different tissues extracted from individuals at three different stages of development. We thus applied three linear models at larval, pre-pupal and adult stages using the \u003cem\u003elm\u003c/em\u003e function from the \u003cem\u003estats\u003c/em\u003e package, with either untransformed or square-root transformed cadmium concentration as the response variable, the tissue, the treatment, and the interaction between tissue and treatment as explanatory variables. We then conducted and analysis of variance using the \u003cem\u003eAnova\u003c/em\u003e function from the \u003cem\u003ecar\u003c/em\u003e package. Post-hoc tests analyses were performed using the \u003cem\u003elsmeans\u003c/em\u003e function from the \u003cem\u003eemmeans\u003c/em\u003e package. As there was no evidence of a difference in cadmium concentration between males and females at any stage (all p\u0026thinsp;\u0026gt;\u0026thinsp;0.10), the sex of individuals was not included in the analysis.\u003c/p\u003e \u003cp\u003eFor linear-mixed effect model with interaction terms, omnibus analyses of variance were conducted using the Kenward-Roger approximation for the computation of fixed effects degrees of freedom using the R package \u003cem\u003elmerTest\u003c/em\u003e (Kuznetsova et al., \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2017\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eModelling assumptions (normality of residuals, normality of random effects, residuals linearity, homogeneity of variance, collinearity of factors) were checked by visual inspection of the residuals using the \u003cem\u003echeck_model\u003c/em\u003e function of the \u003cem\u003eperformance\u003c/em\u003e package (L\u0026uuml;decke et al., \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Following recommendations of the American Statistical Association (Wasserstein and Lazar, \u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e2016\u003c/span\u003e), we do not use an arbitrary threshold (e.g. 0.05) to interpret our results, but instead we use p-values to assess the strength of the statistical evidence to reject the null hypothesis.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e"},{"header":"3. Results","content":"\u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003e3.1. Effect of treatment on food consumption, body mass, relative growth rate and larval mortality\u003c/h2\u003e \u003cp\u003eSummary statistics (sample size, median, mean\u0026thinsp;\u0026plusmn;\u0026thinsp;sd, min and maximum values) for food consumption, mass and relative growth rate across developmental stages and treatments are presented in Table \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e \u0026ndash; S3. To explore the relationship between cadmium treatments and individual food consumption or larval mass, our models were designed to compare changes in these variables over six days for each treatment. The inclusion of a quadratic term proved to be the most suitable approach, particularly due to the distribution of the data and marked changes in food consumption or mass observed between day 0 and day 6. Food intake on day 0 was 100% for all treatments and there was no difference in mass at day 0 between cadmium the treated subjects and the controls (p\u0026thinsp;=\u0026thinsp;0.71). In addition, the food consumption of larvae from the highest treatment T9 on day 6 could not be monitored as all larvae died before data collection.\u003c/p\u003e \u003cp\u003eWe observed small to strong statistical evidence suggesting that larvae from treatments T3 (31.25 \u0026micro;g) and from T5 (125 \u0026micro;g) to T9 (50 mg) exhibited higher food consumption on day 1 compared to the control group (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). In addition, T3 to T9 treatments, including T4 (62.50 \u0026micro;g), showed a distinct pattern from day 1 to day 4, with food consumption consistently higher than that of the control group. From day 4 to day 6, this pattern of increased food consumption became even more pronounced in these treatments than in the controls. This trend correlates with the interaction between day and treatment and the linear component of the quadratic term model (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eSummary of the linear mixed model investigating the effect of treatments on food consumption (logit-transformed) over six days, fitted using a restricted maximum likelihood function. Model includes continuous variable days, treatment, the interaction between days and treatment as well as quadratic terms. The table shows model estimates\u0026thinsp;\u0026plusmn;\u0026thinsp;standard error (Est. \u0026plusmn; s.e.), associated 95% confidence intervals (C.I.) and p-values.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFixed effects\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eEst. \u0026plusmn; s.e.\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC.I.\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eP\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIntercept (day 1, control)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e3.45\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[3.37\u0026ndash;3.53]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDay\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.07\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[0.00\u0026ndash;0.13]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.034\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTreatment\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT1\u0026ndash;7.80 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[-0.11\u0026ndash;0.11]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.969\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT2\u0026ndash;15.625 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-0.03\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[-0.14\u0026ndash;0.08]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.57\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT3\u0026ndash;31.25 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.11\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[0.00\u0026ndash;0.22]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.045\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT4\u0026ndash;62.50 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.09\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[-0.02\u0026ndash;0.20]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.102\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT5\u0026ndash;125 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.13\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[0.02\u0026ndash;0.24]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.025\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT6\u0026ndash;250 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.14\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[0.03\u0026ndash;0.25]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.015\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT7\u0026ndash;0.5 mg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.17\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[0.06\u0026ndash;0.29]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.002\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT8\u0026ndash;5 mg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.15\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[0.04\u0026ndash;0.26]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.009\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT9\u0026ndash;50 mg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.19\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[0.07\u0026ndash;0.30]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.002\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDay\u003csup\u003e2\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-0.04\u0026thinsp;\u0026plusmn;\u0026thinsp;0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[-0.05\u0026nbsp;\u0026ndash;\u0026nbsp;-0.03]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDay \u0026times; Treatment\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT1\u0026ndash;7.80 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-0.03\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[-0.12\u0026ndash;0.05]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.465\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT2\u0026ndash;15.625 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.02\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[-0.07\u0026ndash;0.11]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.652\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT3\u0026ndash;31.25 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-0.09\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[-0.18\u0026nbsp;\u0026ndash;\u0026nbsp;-0.01]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.032\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT4\u0026ndash;62.50 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-0.09\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[-0.18\u0026nbsp;\u0026ndash;\u0026nbsp;-0.01]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.034\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT5\u0026ndash;125 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-0.09\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[-0.17\u0026nbsp;\u0026ndash;\u0026nbsp;-0.00]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.048\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT6\u0026ndash;250 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-0.12\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[-0.20\u0026nbsp;\u0026ndash;\u0026nbsp;-0.03]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.008\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT7\u0026ndash;0.5 mg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-0.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[-0.29\u0026nbsp;\u0026ndash;\u0026nbsp;-0.12]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT8\u0026ndash;5 mg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-0.07\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[-0.16\u0026ndash;0.01]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.092\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT9\u0026ndash;50 mg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-0.12\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[-0.24\u0026nbsp;\u0026ndash;\u0026nbsp;-0.00]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.044\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDay\u003csup\u003e2\u003c/sup\u003e \u0026times; Treatment\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT1\u0026ndash;7.80 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.01\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[-0.00\u0026ndash;0.03]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.096\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT2\u0026ndash;15.625 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[-0.01\u0026ndash;0.01]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.962\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT3\u0026ndash;31.25 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.01\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[0.00\u0026ndash;0.03]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.037\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT4\u0026ndash;62.50 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.02\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[0.00\u0026ndash;0.03]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.017\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT5\u0026ndash;125 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.02\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[0.00\u0026ndash;0.03]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.008\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT6\u0026ndash;250 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.03\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[0.02\u0026ndash;0.05]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT7\u0026ndash;0.5 mg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.06\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[0.05\u0026ndash;0.08]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT8\u0026ndash;5 mg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.03\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[0.02\u0026ndash;0.05]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT9\u0026ndash;50 mg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.05\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[0.02\u0026ndash;0.07]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eSimilarly, the difference in mass between the larvae treated with higher doses (T6 to T9) and the control group increased over time. Larvae from treatments T6 and T7 showed a lower mass on day 1 compared to the control group, with this difference decreasing over time (i.e. individuals in this group compensate for their delayed mass gain compared to the control group). However, for treatments T8 and T9, the mass remains lower throughout the study period, indicating a sustained negative impact of these highest doses on larval mass (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e, Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eSummary of the linear mixed model investigating the effect of treatment on mass (log-transformed) over six days fitted using a restricted maximum likelihood function. Model includes continuous variable days, treatment, the interaction between days and treatment as well as quadratic terms. The table shows model estimates\u0026thinsp;\u0026plusmn;\u0026thinsp;standard error (Est. \u0026plusmn; s.e.) and associated 95% confidence intervals (C.I.); for log-transformed variables, the table also shows multiplicative estimates for untransformed variable (Est. mult.) and associated multiplicative confidence intervals (C.I. mult.); p-values. The multiplicative estimate of the intercept corresponds to the geometric mean of the variable.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFixed effects\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eEst. \u0026plusmn; s.e.\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC.I.\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eEst. Mult.\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eC.I. mult.\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eP\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIntercept (day 0, control)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-3.12\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-3.25\u0026nbsp;\u0026ndash;\u0026nbsp;-2.99]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e[0.04\u0026ndash;0.05]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDay\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.56\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[0.46\u0026ndash;0.66]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1.75\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e[1.59\u0026ndash;1.93]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTreatment\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT1\u0026ndash;7.80 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-0.18\u0026ndash;0.17]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e[0.83\u0026ndash;1.19]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.957\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT2\u0026ndash;15.625 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-0.02\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-0.20\u0026ndash;0.16]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.98\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e[0.82\u0026ndash;1.17]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.798\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT3\u0026ndash;31.25 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-0.07\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-0.25\u0026ndash;0.11]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.93\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e[0.78\u0026ndash;1.11]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.424\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT4\u0026ndash;62.50 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-0.07\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-0.25\u0026ndash;0.11]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.93\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e[0.78\u0026ndash;1.12]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.454\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT5\u0026ndash;125 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-0.14\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-0.32\u0026ndash;0.04]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.87\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e[0.73\u0026ndash;1.04]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.132\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT6\u0026ndash;250 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-0.06\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-0.24\u0026ndash;0.12]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.94\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e[0.79\u0026ndash;1.12]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.505\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT7\u0026ndash;0.5 mg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-0.07\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-0.25\u0026ndash;0.11]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.93\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e[0.78\u0026ndash;1.11]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.422\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT8\u0026ndash;5 mg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.07\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-0.11\u0026ndash;0.25]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1.08\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e[0.90\u0026ndash;1.29]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.425\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT9\u0026ndash;50 mg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.09\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-0.09\u0026ndash;0.27]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1.09\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e[0.91\u0026ndash;1.31]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.329\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDay\u003csup\u003e2\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-0.01\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-0.03\u0026ndash;0.00]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.99\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e[0.97\u0026ndash;1.00]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.096\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDay \u0026times; Treatment\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT1\u0026ndash;7.80 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.05\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-0.09\u0026ndash;0.19]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1.05\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e[0.91\u0026ndash;1.21]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.501\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT2\u0026ndash;15.625 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.01\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-0.13\u0026ndash;0.15]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1.01\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e[0.88\u0026ndash;1.16]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.873\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT3\u0026ndash;31.25 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.02\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-0.12\u0026ndash;0.16]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1.02\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e[0.89\u0026ndash;1.18]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.759\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT4\u0026ndash;62.50 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-0.05\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-0.19\u0026ndash;0.09]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.95\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e[0.83\u0026ndash;1.09]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.464\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT5\u0026ndash;125 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-0.1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-0.24\u0026ndash;0.04]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.9\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e[0.79\u0026ndash;1.04]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.153\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT6\u0026ndash;250 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-0.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-0.34\u0026nbsp;\u0026ndash;\u0026nbsp;-0.06]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.82\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e[0.71\u0026ndash;0.94]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.004\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT7\u0026ndash;0.5 mg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-0.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-0.34\u0026nbsp;\u0026ndash;\u0026nbsp;-0.06]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.82\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e[0.71\u0026ndash;0.94]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.005\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT8\u0026ndash;5 mg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-0.68\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-0.82\u0026nbsp;\u0026ndash;\u0026nbsp;-0.54]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.5\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e[0.44\u0026ndash;0.58]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT9\u0026ndash;50 mg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-0.71\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-0.85\u0026nbsp;\u0026ndash;\u0026nbsp;-0.57]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.49\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e[0.43\u0026ndash;0.56]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDay\u003csup\u003e2\u003c/sup\u003e \u0026times; Treatment\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT1\u0026ndash;7.80 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-0.01\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-0.03\u0026ndash;0.01]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.99\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e[0.97\u0026ndash;1.01]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.341\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT2\u0026ndash;15.625 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-0.02\u0026ndash;0.02]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e[0.98\u0026ndash;1.02]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.859\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT3\u0026ndash;31.25 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-0.02\u0026ndash;0.02]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e[0.98\u0026ndash;1.02]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.954\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT4\u0026ndash;62.50 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.01\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-0.01\u0026ndash;0.03]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1.01\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e[0.99\u0026ndash;1.03]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.419\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT5\u0026ndash;125 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.01\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-0.01\u0026ndash;0.04]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1.01\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e[0.99\u0026ndash;1.04]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.245\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT6\u0026ndash;250 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.01\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-0.01\u0026ndash;0.04]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1.01\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e[0.99\u0026ndash;1.04]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.218\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT7\u0026ndash;0.5 mg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-0.02\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-0.04\u0026ndash;0.00]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.98\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e[0.96\u0026ndash;1.00]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.116\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT8\u0026ndash;5 mg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.04\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[0.01\u0026ndash;0.06]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1.04\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e[1.01\u0026ndash;1.06]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT9\u0026ndash;50 mg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.02\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[0.00\u0026ndash;0.05]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1.02\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e[1.00\u0026ndash;1.05]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.039\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eComparison of the relative growth rate (RGR) between treatments confirms the results previously described, with strong statistical evidence for a decrease in RGR from treatment T5 to T9 (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e, Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eSummary of the linear mixed model investigating the effect of treatment on Relative Growth Rate fitted using a restricted maximum likelihood function. Model includes treatment only. The table shows model estimates\u0026thinsp;\u0026plusmn;\u0026thinsp;standard error (Est. \u0026plusmn; s.e.) and associated 95% confidence intervals (C.I.).\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFixed effects\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eEst. \u0026plusmn; s.e.\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC.i.\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003ep\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIntercept (control)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e3.37\u0026thinsp;\u0026plusmn;\u0026thinsp;0.24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[2.90\u0026ndash;3.85]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTreatment\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT1\u0026ndash;7.80 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-0.31\u0026thinsp;\u0026plusmn;\u0026thinsp;0.34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[-0.98\u0026ndash;0.37]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.372\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT2\u0026ndash;15.625 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-0.18\u0026thinsp;\u0026plusmn;\u0026thinsp;0.34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[-0.86\u0026ndash;0.49]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.59\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT3\u0026ndash;31.25 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.08\u0026thinsp;\u0026plusmn;\u0026thinsp;0.34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[-0.59\u0026ndash;0.76]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.808\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT4\u0026ndash;62.50 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-0.29\u0026thinsp;\u0026plusmn;\u0026thinsp;0.34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[-0.97\u0026ndash;0.38]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.393\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT5\u0026ndash;125 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-0.91\u0026thinsp;\u0026plusmn;\u0026thinsp;0.34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[-1.59\u0026nbsp;\u0026ndash;\u0026nbsp;-0.24]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.008\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT6\u0026ndash;250 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-2.05\u0026thinsp;\u0026plusmn;\u0026thinsp;0.34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[-2.72\u0026nbsp;\u0026ndash;\u0026nbsp;-1.38]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT7\u0026ndash;0.5 mg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-3.09\u0026thinsp;\u0026plusmn;\u0026thinsp;0.34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[-3.77\u0026nbsp;\u0026ndash;\u0026nbsp;-2.42]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT8\u0026ndash;5 mg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-3.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[-4.00\u0026nbsp;\u0026ndash;\u0026nbsp;-2.66]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT9\u0026ndash;50 mg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-3.38\u0026thinsp;\u0026plusmn;\u0026thinsp;0.35\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e[-4.07\u0026nbsp;\u0026ndash;\u0026nbsp;-2.69]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eLastly, the larval mortality of each treatment revealed a lethal dose threshold, since 100% of the larvae died in T7, T8 and T9 treatments, while no difference in mortality rates was found between the other treatments and the control group (all p\u0026thinsp;\u0026gt;\u0026thinsp;0.10; Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003e3.2. Bioaccumulation of cadmium across developmental stages and tissues\u003c/h2\u003e \u003cp\u003eFor both T2 (15.625 \u0026micro;g) and T5 (125 \u0026micro;g) treatments, cadmium assays demonstrate dose-dependent bioaccumulation during the post-embryonic development compared to the control group (Table\u0026nbsp;4, Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e), with variations according to stages and treatment. Notably, the T2 subjects experienced minor fluctuations compared to those exposed to the highest dose, with a peak at 9-day in the pupal stage. Interestingly, cadmium levels in T5 adults were similar to those of the 4th larval stage, indicating a decrease as the larvae matured into adults (Table\u0026nbsp;4 and Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"No\" id=\"Taba\" border=\"1\"\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colspan=\"6\" nameend=\"c6\" namest=\"c1\"\u003e \u003cp\u003eTable\u0026nbsp;4: Summary of the linear mixed model investigating the effect of Spodoptera littoralis developmental stages and treatment on log-transformed cadmium concentration in individuals using a restricted maximum likelihood function. Model includes developmental stages, treatment and the interaction between stages and treatment. The table shows model estimates\u0026thinsp;\u0026plusmn;\u0026thinsp;standard error (Est. \u0026plusmn; s.e.) and associated 95% confidence intervals (C.I.); for log-transformed variables, the table also shows multiplicative estimates for untransformed variable (Est. mult.) and associated multiplicative confidence intervals (C.I. mult.); p-values. The multiplicative estimate of the intercept corresponds to the geometric mean of the variable.\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFixed effects\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003eEst. \u0026plusmn; s.e.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003eC.I.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003eEst. Mult.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003eC.I. mult.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003eP\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIntercept (L4, control)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-4.61\u0026thinsp;\u0026plusmn;\u0026thinsp;0.32\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-5.25\u0026nbsp;\u0026ndash;\u0026nbsp;-3.97]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e[0.01\u0026ndash;0.02]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eStage\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eL5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.57\u0026thinsp;\u0026plusmn;\u0026thinsp;0.38\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[0.82\u0026ndash;2.32]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e4.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e[2.27\u0026ndash;10.14]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eL6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.05\u0026thinsp;\u0026plusmn;\u0026thinsp;0.36\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-0.66\u0026ndash;0.76]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e[0.52\u0026ndash;2.14]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.89\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePre-pupae\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.07\u0026thinsp;\u0026plusmn;\u0026thinsp;0.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-0.67\u0026ndash;0.81]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e[0.51\u0026ndash;2.24]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.85\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3-days pupae\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.14\u0026thinsp;\u0026plusmn;\u0026thinsp;0.40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-0.65\u0026ndash;0.92]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e[0.52\u0026ndash;2.50]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.73\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e9-days pupae\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-0.03\u0026thinsp;\u0026plusmn;\u0026thinsp;0.40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-0.81\u0026ndash;0.76]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.97\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e[0.45\u0026ndash;2.13]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.95\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAdult\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.01\u0026thinsp;\u0026plusmn;\u0026thinsp;0.40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-0.77\u0026ndash;0.79]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e[0.46\u0026ndash;2.21]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.98\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTreatment\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e15.625 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e6.34\u0026thinsp;\u0026plusmn;\u0026thinsp;0.40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[5.55\u0026ndash;7.12]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e564.53\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e[258.39\u0026ndash;1233.39]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e125 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e8.26\u0026thinsp;\u0026plusmn;\u0026thinsp;0.40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[7.47\u0026ndash;9.04]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3846.81\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e[1760.70\u0026ndash;8404.59]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eStage \u0026times; Treatment\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eStage L5 \u0026times; 15.625 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-1.32\u0026thinsp;\u0026plusmn;\u0026thinsp;0.48\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-2.27\u0026nbsp;\u0026ndash;\u0026nbsp;-0.37]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e[0.10\u0026ndash;0.69]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.01\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eStage L6 \u0026times; 15.625 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.26\u0026thinsp;\u0026plusmn;\u0026thinsp;0.46\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-0.65\u0026ndash;1.18]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e[0.52\u0026ndash;3.25]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.57\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eStage pre-pupae \u0026times; 15.625 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.80\u0026thinsp;\u0026plusmn;\u0026thinsp;0.47\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-0.14\u0026ndash;1.74]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e[0.87\u0026ndash;5.69]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.09\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eStage 3-days pupae \u0026times; 15.625 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.51\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-0.01\u0026ndash;2.01]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.72\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e[0.99\u0026ndash;7.47]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.05\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eStage 9-days pupae \u0026times; 15.625 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.18\u0026thinsp;\u0026plusmn;\u0026thinsp;0.51\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[0.17\u0026ndash;2.19]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e[1.18\u0026ndash;8.91]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.02\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eStage adult \u0026times; 15.625 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.01\u0026thinsp;\u0026plusmn;\u0026thinsp;0.54\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-0.05\u0026ndash;2.07]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.75\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e[0.95\u0026ndash;7.92]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.06\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eStage L5 \u0026times; 125 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-1.24\u0026thinsp;\u0026plusmn;\u0026thinsp;0.46\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-2.15\u0026nbsp;\u0026ndash;\u0026nbsp;-0.32]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e[0.12\u0026ndash;0.72]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.01\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eStage L6 \u0026times; 125 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-0.43\u0026thinsp;\u0026plusmn;\u0026thinsp;0.47\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-1.35\u0026ndash;0.50]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.65\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e[0.26\u0026ndash;1.65]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.36\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eStage pre-pupae \u0026times; 125 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.61\u0026thinsp;\u0026plusmn;\u0026thinsp;0.50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-0.39\u0026ndash;1.60]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.84\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e[0.68\u0026ndash;4.97]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.23\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eStage 3-days pupae \u0026times; 125 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.73\u0026thinsp;\u0026plusmn;\u0026thinsp;0.50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-0.25\u0026ndash;1.72]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e[0.78\u0026ndash;5.58]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.14\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eStage 9-days pupae \u0026times; 125 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.95\u0026thinsp;\u0026plusmn;\u0026thinsp;0.51\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-0.06\u0026ndash;1.96]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e[0.95\u0026ndash;7.12]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.06\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eStage adult \u0026times; 125 \u0026micro;g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.22\u0026thinsp;\u0026plusmn;\u0026thinsp;0.51\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e[-0.79\u0026ndash;1.22]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e[0.45\u0026ndash;3.40]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.67\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eTo go further, cadmium concentrations after T4 and T5 treatments were measured in various tissues of the final larval stage, pre-pupa and adult (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). We found a difference in cadmium concentration between treatment and the interaction between tissue-treatment for all the three stages (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e5\u003c/span\u003e). In the larval control group, a low amount of cadmium was detected in the digestive tract and in the head, likely due to a slight contamination during the measurement process. The other control samples showed levels below the detection limit. Furthermore, post-hoc tests showed no statistical difference in cadmium concentration between the different tissues of the control group at any stage. In contrast, T5-treated larvae exhibited accumulation of cadmium in their digestive tract and fat body. The tissues with the highest bioaccumulation are not the same according to treatment. For example, during the pre-pupal stage, the higher concentrations were observed in the feces of the T5 group but in the exuviae of the T4 (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). The group exposed to the highest level of cadmium showed the most substantial accumulation in most tissues, except for the integument. In addition, cadmium levels in the feces of pre-pupae reached up to four times the ingested dose and between 10 to 25% of the cadmium was found in the integument, but we did not report any difference in concentration between these two matrices (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). In T4 and T5-treated adults, cadmium predominantly concentrated in the abdomen (average of 59.98 \u0026micro;g/g and 32.97 \u0026micro;g/g, respectively). Cadmium concentrations were barely detectable in T4-adult antennae and haemolymph (average of 0.40 \u0026micro;g/g and 0.04 \u0026micro;g/g, respectively) and slightly higher in T5 adults (average of 2.53 \u0026micro;g/g and 1.56 \u0026micro;g/g, respectively).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 5\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eAnalysis of variance testing the difference in cadmium concentrations in different tissues and treatment using Kenward-Roger approximation for the computation of fixed effects degrees of freedom. The table shows the sum of squares, degrees of freedom (Df), F and P values.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colspan=\"4\" nameend=\"c5\" namest=\"c2\"\u003e \u003cp\u003eLarvae\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFixed effects\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003eSum of squares\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003eDf\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003eF\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003eP\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTissues\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e17.275\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e75.639\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTreatment\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e248.976\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1090.171\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTissues \u0026times; Treatment\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e50.791\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e111.196\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colspan=\"4\" nameend=\"c5\" namest=\"c2\"\u003e \u003cp\u003e\u003cb\u003ePre-pupae\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFixed effects\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003eSum of squares\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003eDf\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003eF\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003eP\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTissues\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e248.36\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e190.83\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTreatment\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1546.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1188.26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTissues \u0026times; Treatment\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e347.79\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e133.61\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colspan=\"4\" nameend=\"c5\" namest=\"c2\"\u003e \u003cp\u003e\u003cb\u003eAdults\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFixed effects\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003eSum of squares\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003eDf\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003eF\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003eP\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTissues\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e10967.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e66.63\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTreatment\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1985.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e24.121\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTissues \u0026times; Treatment\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e7074.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e21.49\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"4. Discussion","content":"\u003cp\u003eIn this study, we explored the impact of ingested cadmium at sublethal to lethal doses on several life history traits of the model crop pest \u003cem\u003eSpodoptera littoralis\u003c/em\u003e, including food intake by larvae, post-embryonic development and bioaccumulation. Our results indicate that sublethal doses of cadmium significantly affect food consumption and relative growth rate in larvae over a 6-day period, and to a lesser extent affect their mass after treatments at higher doses. The adverse effects become pronounced at a relatively low-dose of 31.25 µg/g, with particularly severe outcomes at the highest – lethal – doses ranging from 0.5 to 50 mg/g, where 100% mortality was observed post-treatment. Additionally, while higher cadmium treatments starting from 250 µg/g impacted larval mass, we observed that by day 6 before metamorphosis, larvae receiving sublethal doses had regained mass comparable to the control group, suggesting a selective pressure that allows individuals with initially lower mass to compensate for their delayed growth despite reduced food intake. Contrasting findings have been reported in other studies regarding the impact of cadmium on mass, where a decrease in larval body mass was noted under sublethal cadmium exposure, but not always with a recovery to control levels (Humann-Guilleminot et al., \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Vlahović et al., \u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e2017\u003c/span\u003e, \u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). The variation in larval mass may stem from the species' ability to detoxify, a process that can vary in energy consumption (Castañeda et al., \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2009\u003c/span\u003e; Vlahović et al., \u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Specifically, \u003cem\u003eSpodoptera littoralis\u003c/em\u003e is believed to possess a high capacity for detoxification (Aviles et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2019\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe dose-dependent relationship observed after the treatments in the present study is consistent with the growth reductions previously reported in this moth (Humann-Guilleminot et al., \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2024\u003c/span\u003e) and in other species belonging to different insects orders (e.g., \u003cem\u003eAiolopus thalassinus\u003c/em\u003e, \u003cem\u003eOncopeltus fasciatus\u003c/em\u003e, \u003cem\u003eLymantria dispar\u003c/em\u003e, \u003cem\u003eChironomus riparius\u003c/em\u003e, \u003cem\u003ePoecilus cupreus\u003c/em\u003e) chronically exposed to heavy metals (Cervera et al., \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2004\u003c/span\u003e; Maryanski et al., \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Ortel, \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e1995\u003c/span\u003e; Schmidt et al., \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e1992\u003c/span\u003e; Sildanchandra and Crane, \u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e2000\u003c/span\u003e). Some studies reported a potential behavioural adaptation of larvae to avoid cadmium contaminated food, where larvae decrease food consumption to ensure survival, thus reducing the intake of cadmium – but also essential nutrients – at the expense of its growth (Migula and Binkowska, \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e1993\u003c/span\u003e; van Straalen and Hoffmann, \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e2000\u003c/span\u003e). Indeed, some insects have the ability to detect contaminants in their food, leading them to adjust their consumption, which in turn results in lower mass (Nummelin et al., \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2007\u003c/span\u003e; van Capelleveen et al., \u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e1986\u003c/span\u003e; Vlahović et al., \u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). These findings are similar to \u003cem\u003eDrosophila melanogaster\u003c/em\u003e that showed an aversion to cadmium-contaminated food (Bahadorani and Hilliker, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2009\u003c/span\u003e), although this adaptive strategy would be viable only on the short term. While we observed differences in food consumption between treated and control groups, these differences were unexpected as treated larvae consumed more than the controls at the approach of the pre-pupal stage. As observed in the control group, healthy individuals generally consume less when preparing for metamorphosis, often by purging just before forming the chrysalis (Kraus et al., \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). However, in instances where development is delayed, as it has been shown with cadmium (Humann-Guilleminot et al., \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2024\u003c/span\u003e), this reduction in food consumption may also occurs later than usual. Interestingly, the increased food consumption observed in the treated groups did not translate into mass gain, suggesting a physiological cost associated with the body's detoxification processes. Another possible explanation for this phenomenon is that ingested cadmium is predominantly stored in the insects' guts, which may impact their digestive enzymes, affecting nutrient absorption (Hensbergen et al., \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2000\u003c/span\u003e; Su et al., \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Zhang et al., \u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e2020\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAt the larval stage, we observed a trend toward cadmium accumulation, reaching a saturation threshold in the pupal stage, followed by a decrease in the adult stage. These results are consistent with observations in \u003cem\u003eLymantria dispar\u003c/em\u003e, where cadmium levels continuously increase during larval development, peaking at the pupal stage, and then decreasing in adults (Gintenreiter et al., \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e1993\u003c/span\u003e). This suggests that a significant amount of cadmium was eliminated during metamorphosis, particularly after exposure to higher levels. It has been shown under these conditions that metals are purged primarily during this phase, resulting in concentrations in adults up to 125 times lower than those in larvae (Kraus et al., \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Lindqvist and Block, 1995). Our results show that insects excrete a significant amount of cadmium through the integument and exuviae during metamorphosis, and do not absorb all ingested cadmium as evidenced by high concentrations in their feces. Numerous studies on insects from various orders and habitats have shown that cadmium accumulates mainly in their digestive tract at larval stage, serving as a major organ of protection against metals, by trapping them in epithelial cells (Aoki et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e1984\u003c/span\u003e; Dallinger, \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e1993\u003c/span\u003e; Kafel et al., \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2003\u003c/span\u003e). However, the cadmium absorbed by the midgut is then transported by the haemolymph and distributed in all parts of the body (Kafel et al., \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2003\u003c/span\u003e; Leonard et al., \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2009\u003c/span\u003e) which probably explains the concentrations measured here in the heads of larvae and adults, and in the antennae. In addition, cadmium stored in the meconium – a metabolic waste– has been shown to be excreted upon adult emergence (Aoki and Suzuki, \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e1984\u003c/span\u003e; Dallinger, \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e1993\u003c/span\u003e; Vlahović et al., \u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e2017\u003c/span\u003e), which could explain the significant reduction in cadmium levels observed in \u003cem\u003eS. littoralis\u003c/em\u003e adults compared to the pre-pupal stage (Dallinger, \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e1993\u003c/span\u003e; Dar et al., \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Lindqvist and Block, 1995). Altogether, these results highlight the critical role of cadmium excretion processes during the larval stages (feces, integument), metamorphosis (exuviae) and in adults in \u003cem\u003eS. littoralis\u003c/em\u003e, and in insects in general.\u003c/p\u003e "},{"header":"5. Conclusion","content":"\u003cp\u003eWe have demonstrated the effects of nine cadmium concentrations on food consumption and larval growth in the noctuid moth \u003cem\u003eSpodoptera littoralis\u003c/em\u003e. Our results show that cadmium concentrations peak during the pupal stage and then decrease in adults, indicating effective detoxification mechanisms during metamorphosis. This is confirmed by the significant amounts of cadmium excreted in feces, integument, and exuviae at the prepupal stage. In addition, we observe that larvae stored most of the ingested cadmium in their digestive tract, from where it was transferred by haemolymph to various body parts, including the head and the antennae. These results confirm the toxicity of cadmium and its role as an environmental stress factor. We recommend further research on the chronic effects of cadmium, with particular emphasis on the impact of contamination by sublethal doses throughout the food chain, at different trophic levels, considering potential synergistic impacts with other environmental pollutant.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cu\u003eEthical statement:\u0026nbsp;\u003c/u\u003eWe hereby confirm that this material is the authors\u0026apos; original work, has not been previously published, and accurately reflects their own research and analysis. The paper credits the significant contributions of co-authors and collaborators, and the results are properly contextualized within prior and existing research. All authors have been actively involved in the substantial work leading to this paper and take full public responsibility for its content.\u003c/p\u003e\n\u003cp\u003e\u003cu\u003eFunding:\u003c/u\u003e The project was supported by an Emergence funding program of Sorbonne University.\u003c/p\u003e\n\u003cp\u003e\u003cu\u003eAuthors\u0026rsquo; contributions\u003c/u\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eS\u0026eacute;gol\u0026egrave;ne Humann-Guilleminot:\u0026nbsp;\u003c/strong\u003eData curation, Formal analysis, Visualization, Writing \u0026ndash; original draft, Writing \u0026ndash; review \u0026amp; editing.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMarie-Anne Pottier:\u003c/strong\u003e Conceptualization, Investigation, Methodology.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMaryse Rouelle:\u003c/strong\u003e Investigation,\u0026nbsp;Writing \u0026ndash; review \u0026amp; editing.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAnnick Maria:\u003c/strong\u003e Methodology, Resource.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMartine Maibeche:\u003c/strong\u003e Conceptualization, Funding acquisition, Project administration, Supervision, Writing \u0026ndash; review \u0026amp; editing.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDavid Siaussat:\u003c/strong\u003e Conceptualization, Funding acquisition, Project administration, Supervision, Writing \u0026ndash; review \u0026amp; editing.\u003c/p\u003e\u003ch2\u003eData Availability\u003c/h2\u003e\u003cp\u003eAll data used in this study is included within its Supplementary Information files.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAbedi, T., Mojiri, A., 2020. Cadmium uptake by wheat (Triticum aestivum L.): An overview. Plants 9. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/plants9040500\u003c/span\u003e\u003cspan address=\"10.3390/plants9040500\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAli, H., Khan, E., 2018. Bioaccumulation of non-essential hazardous heavy metals and metalloids in freshwater fish. Risk to human health. Environmental Chemistry Letters 16, 903\u0026ndash;917. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10311-018-0734-7\u003c/span\u003e\u003cspan address=\"10.1007/s10311-018-0734-7\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAli, H., Khan, E., Ilahi, I., 2019. Environmental chemistry and ecotoxicology of hazardous heavy metals: environmental persistence, toxicity, and bioaccumulation. Journal of Chemistry 2019, 1\u0026ndash;301. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1155/2019/6730305\u003c/span\u003e\u003cspan address=\"10.1155/2019/6730305\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAoki, Y., Suzuki, K.T., 1984. Excretion of cadmium and change in the relative ratio of iso-cadmium-binding proteins during metamorphosis of fleshfly (Sarcophaga peregrina). Comparative Biochemistry and Physiology Part C: Comparative Pharmacology 78, 315\u0026ndash;317. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/0742-8413(84)90089-6\u003c/span\u003e\u003cspan address=\"10.1016/0742-8413(84)90089-6\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAoki, Y., Suzuki, K.T., Kubota, K., 1984. Accumulation of cadmium and induction of its binding protein in the digestive tract of fleshfly (Sarcophaga peregrina) larvae. Comparative Biochemistry and Physiology Part C: Comparative Pharmacology 77, 279\u0026ndash;282. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/0742-8413(84)90013-6\u003c/span\u003e\u003cspan address=\"10.1016/0742-8413(84)90013-6\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAviles, A., Boulogne, I., Durand, N., Maria, A., Cordeiro, A., Bozzolan, F., Goutte, A., Alliot, F., Dacher, M., Renault, D., Maibeche, M., Siaussat, D., 2019. Effects of DEHP on post-embryonic development, nuclear receptor expression, metabolite and ecdysteroid concentrations of the moth Spodoptera littoralis. Chemosphere 215, 725\u0026ndash;738. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.chemosphere.2018.10.102\u003c/span\u003e\u003cspan address=\"10.1016/j.chemosphere.2018.10.102\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBahadorani, S., Hilliker, A.J., 2009. Biological and behavioral effects of heavy metals in Drosophila melanogaster adults and larvae. Journal of Insect Behavior 22, 399\u0026ndash;411. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10905-009-9181-4\u003c/span\u003e\u003cspan address=\"10.1007/s10905-009-9181-4\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBradl, H.B., 2005. Chapter 1 Sources and origins of heavy metals, in: Bradl, H.B. (Ed.), Interface Science and Technology. Elsevier, pp. 1\u0026ndash;27. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/S1573-4285(05)80020-1\u003c/span\u003e\u003cspan address=\"10.1016/S1573-4285(05)80020-1\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBrown, E.S., Dewhurst, C.F., 1975. The genus Spodoptera (Lepidoptera, Noctuidae) in Africa and the near East. Bulletin of Entomological Research 65, 221\u0026ndash;262. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1017/S0007485300005939\u003c/span\u003e\u003cspan address=\"10.1017/S0007485300005939\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBrunn, H., Arnold, G., K\u0026ouml;rner, W., Rippen, G., Steinh\u0026auml;user, K.G., Valentin, I., 2023. PFAS: forever chemicals\u0026mdash;persistent, bioaccumulative and mobile. Reviewing the status and the need for their phase out and remediation of contaminated sites. Environmental Sciences Europe 35, 20. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1186/s12302-023-00721-8\u003c/span\u003e\u003cspan address=\"10.1186/s12302-023-00721-8\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eButt, A., Qurat ul, A., Rehman, K., Khan, M.X., Hesselberg, T., 2018. Bioaccumulation of cadmium, lead, and zinc in agriculture-based insect food chains. Environmental Monitoring and Assessment 190, 698. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10661-018-7051-2\u003c/span\u003e\u003cspan address=\"10.1007/s10661-018-7051-2\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eByers, J.R., Hinks, C.F., 1976. Biosystematics of the genus Euxoa (Lepidoptera: Noctuidae): V. Rearing Procedures, and life cycles of 36 species. The Canadian Entomologist 108, 1345\u0026ndash;1357. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.4039/Ent1081345-12\u003c/span\u003e\u003cspan address=\"10.4039/Ent1081345-12\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCasta\u0026ntilde;eda, L.E., Figueroa, C.C., Fuentes-Contreras, E., Niemeyer, H.M., Nespolo, R.F., 2009. Energetic costs of detoxification systems in herbivores feeding on chemically defended host plants: a correlational study in the grain aphid, Sitobion avenae. The Journal of experimental biology 212, 1185\u0026ndash;90. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1242/jeb.020990\u003c/span\u003e\u003cspan address=\"10.1242/jeb.020990\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCervera, A., Maym\u0026oacute;, A.C., Sendra, M., Mart\u0026iacute;nez-Pardo, R., Garcer\u0026aacute;, M.D., 2004. Cadmium effects on development and reproduction of Oncopeltus fasciatus (Heteroptera: Lygaeidae). Journal of Insect Physiology 50, 737\u0026ndash;749. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.jinsphys.2004.06.001\u003c/span\u003e\u003cspan address=\"10.1016/j.jinsphys.2004.06.001\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eConolly, R.B., Lutz, W.K., 2004. Nonmonotonic dose-response relationships: mechanistic basis, kinetic modeling, and implications for risk assessment. Toxicological Sciences 77, 151\u0026ndash;157. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/toxsci/kfh007\u003c/span\u003e\u003cspan address=\"10.1093/toxsci/kfh007\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDallinger, R., 1993. Strategies of metal detoxification in terestrial invertebrates. Ecotoxicology of Metals in Invertebrates 245\u0026ndash;290.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDar, M.I., Green, I.D., Naikoo, M.I., Khan, F.A., Ansari, A.A., Lone, M.I., 2017. Assessment of biotransfer and bioaccumulation of cadmium, lead and zinc from fly ash amended soil in mustard\u0026ndash;aphid\u0026ndash;beetle food chain. Science of The Total Environment 584\u0026ndash;585, 1221\u0026ndash;1229. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.scitotenv.2017.01.186\u003c/span\u003e\u003cspan address=\"10.1016/j.scitotenv.2017.01.186\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFarrar, R.R., Barbour, J.D., Kennedy, G.G., 1989. Quantifying food consumption and growth in insects. Annals of the Entomological Society of America 82, 593\u0026ndash;598. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/aesa/82.5.593\u003c/span\u003e\u003cspan address=\"10.1093/aesa/82.5.593\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFrat, L., Chertemps, T., Pesce, E., Bozzolan, F., Dacher, M., Planell\u0026oacute;, R., Herrero, O., Llorente, L., Moers, D., Siaussat, D., 2021. Single and mixed exposure to cadmium and mercury in Drosophila melanogaster: Molecular responses and impact on post-embryonic development. Ecotoxicology and Environmental Safety 220, 112377. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.ecoenv.2021.112377\u003c/span\u003e\u003cspan address=\"10.1016/j.ecoenv.2021.112377\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGao, D.-W., Wen, Z.-D., 2016. Phthalate esters in the environment: A critical review of their occurrence, biodegradation, and removal during wastewater treatment processes. Science of The Total Environment 541, 986\u0026ndash;1001. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.scitotenv.2015.09.148\u003c/span\u003e\u003cspan address=\"10.1016/j.scitotenv.2015.09.148\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGintenreiter, S., Ortel, J., Nopp, H.J., 1993. Effects of different dietary levels of cadmium, lead, copper, and zinc on the vitality of the forest pest insect Lymantria dispar L. (Lymantriidae, Lepid). Archives of Environmental Contamination and Toxicology 25, 62\u0026ndash;66. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/BF00230712\u003c/span\u003e\u003cspan address=\"10.1007/BF00230712\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHaider, F.U., Liqun, C., Coulter, J.A., Cheema, S.A., Wu, J., Zhang, R., Wenjun, M., Farooq, M., 2021. Cadmium toxicity in plants: Impacts and remediation strategies. Ecotoxicology and Environmental Safety 211, 111887. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.ecoenv.2020.111887\u003c/span\u003e\u003cspan address=\"10.1016/j.ecoenv.2020.111887\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHealth, E.P. on P., 2015. Scientific Opinion on the pest categorisation of Spodoptera littoralis. EFSA Journal 13, 3987. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.2903/j.efsa.2015.3987\u003c/span\u003e\u003cspan address=\"10.2903/j.efsa.2015.3987\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHensbergen, P.J., van Velzen, M.J.M., Adi Nugroho, R., Donker, M.H., van Straalen, N.M., 2000. Metallothionein-bound cadmium in the gut of the insect Orchesella cincta (Collembola) in relation to dietary cadmium exposure. Comparative Biochemistry and Physiology Part C: Pharmacology, Toxicology and Endocrinology 125, 17\u0026ndash;24. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/S0742-8413(99)00087-0\u003c/span\u003e\u003cspan address=\"10.1016/S0742-8413(99)00087-0\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHumann-Guilleminot, S., Fuentes, A., Maria, A., Couzi, P., Siaussat, D., 2024. Cadmium and phthalate impact developmental growth and mortality of Spodoptera littoralis, but not reproductive success. Ecotoxicology and Environmental Safety 281, 116605. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.ecoenv.2024.116605\u003c/span\u003e\u003cspan address=\"10.1016/j.ecoenv.2024.116605\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eIavicoli, I., Fontana, L., Bergamaschi, A., 2009. The Effects of metals as endocrine disruptors. Journal of Toxicology and Environmental Health, Part B 12, 206\u0026ndash;223. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1080/10937400902902062\u003c/span\u003e\u003cspan address=\"10.1080/10937400902902062\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJin, R., Bu, D., Liu, G., Zheng, M., Lammel, G., Fu, J., Yang, L., Li, C., Habib, A., Yang, Y., Liu, X., 2020. New classes of organic pollutants in the remote continental environment \u0026ndash; Chlorinated and brominated polycyclic aromatic hydrocarbons on the Tibetan Plateau. Environment International 137, 105574. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.envint.2020.105574\u003c/span\u003e\u003cspan address=\"10.1016/j.envint.2020.105574\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKafel, A., Bednarska, K., Augustyniak, M., Witas, I., Szuli\u0026ntilde;ska, E., 2003. Activity of glutathione S-transferase in Spodoptera exigua larvae exposed to cadmium and zinc in two subsequent generations. Environment International 28, 683\u0026ndash;686. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/S0160-4120(02)00111-3\u003c/span\u003e\u003cspan address=\"10.1016/S0160-4120(02)00111-3\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKraus, J.M., Walters, D.M., Wesner, J.S., Stricker, C.A., Schmidt, T.S., Zuellig, R.E., 2014. Metamorphosis Alters Contaminants and Chemical Tracers in Insects: Implications for Food Webs. Environ. Sci. Technol. 48, 10957\u0026ndash;10965. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1021/es502970b\u003c/span\u003e\u003cspan address=\"10.1021/es502970b\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKubier, A., Wilkin, R.T., Pichler, T., 2019. Cadmium in soils and groundwater: A review. Applied Geochemistry 108, 104388. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.apgeochem.2019.104388\u003c/span\u003e\u003cspan address=\"10.1016/j.apgeochem.2019.104388\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKuznetsova, A., Brockhoff, P.B., Christensen, R.H.B., 2017. lmerTest package: tests in linear mixed effects models. Journal of Statistical Software 82, 26. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.18637/jss.v082.i13\u003c/span\u003e\u003cspan address=\"10.18637/jss.v082.i13\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLeonard, E.M., Pierce, L.M., Gillis, P.L., Wood, C.M., O\u0026rsquo;Donnell, M.J., 2009. Cadmium transport by the gut and Malpighian tubules of Chironomus riparius. Aquatic Toxicology 92, 179\u0026ndash;186. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.aquatox.2009.01.011\u003c/span\u003e\u003cspan address=\"10.1016/j.aquatox.2009.01.011\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLindqvist, L., Block, M., 1995. Excretion of cadmium during moulting and metamorphosis in Tenebrio molitor (Coleoptera; Tenebrionidae). Comparative Biochemistry and Physiology Part C: Pharmacology, Toxicology and Endocrinology 111, 325\u0026ndash;328. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/0742-8413(95)00057-U\u003c/span\u003e\u003cspan address=\"10.1016/0742-8413(95)00057-U\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eL\u0026uuml;decke, D., Ben Shachar, M., Patil, I., Waggoner, P., Makowski, D., 2021. Performance: an R package for assessment, comparison and testing of statistical models. The Journal of Open Source Software 6, 3139. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.21105/joss.03139\u003c/span\u003e\u003cspan address=\"10.21105/joss.03139\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMarcus, S.R., Fiumera, A.C., 2016. Atrazine exposure affects longevity, development time and body size in Drosophila melanogaster. Journal of Insect Physiology 91\u0026ndash;92, 18\u0026ndash;25. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.jinsphys.2016.06.006\u003c/span\u003e\u003cspan address=\"10.1016/j.jinsphys.2016.06.006\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMaryanski, M., Kramarz, P., Laskowski, R., Niklinska, M., 2002. Decreased Energetic Reserves, Morphological Changes and Accumulation of Metals in Carabid Beetles (Poecilus cupreus L.) Exposed to Zinc- or Cadmium-contaminated Food. Ecotoxicology 11, 127\u0026ndash;139. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1023/A:1014425113481\u003c/span\u003e\u003cspan address=\"10.1023/A:1014425113481\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMigula, P., Binkowska, K., 1993. Feeding strategies of grasshoppers (Chorthippus sp.) on heavy metal contaminated plants. Science of The Total Environment 134, 1071\u0026ndash;1083. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/S0048-9697(05)80112-3\u003c/span\u003e\u003cspan address=\"10.1016/S0048-9697(05)80112-3\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMoulis, J.-M., 2010. Cellular mechanisms of cadmium toxicity related to the homeostasis of essential metals. BioMetals 23, 877\u0026ndash;896. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10534-010-9336-y\u003c/span\u003e\u003cspan address=\"10.1007/s10534-010-9336-y\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNagorka, R., Koschorreck, J., 2020. Trends for plasticizers in German freshwater environments \u0026ndash; Evidence for the substitution of DEHP with emerging phthalate and non-phthalate alternatives. Environmental Pollution 262, 114237. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.envpol.2020.114237\u003c/span\u003e\u003cspan address=\"10.1016/j.envpol.2020.114237\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNet, S., Semp\u0026eacute;r\u0026eacute;, R., Delmont, A., Paluselli, A., Ouddane, B., 2015. Occurrence, fate, behavior and ecotoxicological state of phthalates in different environmental matrices. Environmental Science \u0026amp; Technology 49, 4019\u0026ndash;4035. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1021/es505233b\u003c/span\u003e\u003cspan address=\"10.1021/es505233b\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNummelin, M., Lodenius, M., Tulisalo, E., Hirvonen, H., Alanko, T., 2007. Predatory insects as bioindicators of heavy metal pollution. Environmental Pollution 145, 339\u0026ndash;347. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.envpol.2006.03.002\u003c/span\u003e\u003cspan address=\"10.1016/j.envpol.2006.03.002\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOrtel, J., 1995. Effects of metals on the total lipid content in the gypsy moth (Lymantria dispar, lymantriidae, lepid.) and its hemolymph. Bulletin of Environmental Contamination and Toxicology 55, 216\u0026ndash;221. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/BF00203012\u003c/span\u003e\u003cspan address=\"10.1007/BF00203012\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePlanell\u0026oacute;, R., Mart\u0026iacute;nez-Guitarte, J.L., Morcillo, G., 2010. Effect of acute exposure to cadmium on the expression of heat-shock and hormone-nuclear receptor genes in the aquatic midge Chironomus riparius. Science of The Total Environment 408, 1598\u0026ndash;1603. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.scitotenv.2010.01.004\u003c/span\u003e\u003cspan address=\"10.1016/j.scitotenv.2010.01.004\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eQuimby, P.C., Frick, K.E., Wauchope, R.D., Kay, S.H., 1979. Effects of cadmium on two biocontrol insects and their host weeds. Bulletin of Environmental Contamination and Toxicology 22, 371\u0026ndash;378. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/BF02026957\u003c/span\u003e\u003cspan address=\"10.1007/BF02026957\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eR Core Team, 2022. R: A language and environment for statistical computing. [WWW Document].\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRaina, R.M., Pawar, P., Sharma, R.N., 2001. Developmental inhibition and reproductive potential impairment in Musca domestica L. by heavy metals. Indian journal of experimental biology 39, 78\u0026ndash;81.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRaise, G., Gemmellaro, M.D., 2024. A review on the effects of heavy metals on the development of blow flies (Diptera: Calliphoridae). WIREs Forensic Science 6, e1503. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1002/wfs2.1503\u003c/span\u003e\u003cspan address=\"10.1002/wfs2.1503\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRana, S.V.S., 2014. Perspectives in endocrine toxicity of heavy metals\u0026mdash;A Review. Biological Trace Element Research 160, 1\u0026ndash;14. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s12011-014-0023-7\u003c/span\u003e\u003cspan address=\"10.1007/s12011-014-0023-7\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRathi, B.S., Kumar, P.S., Vo, D.-V.N., 2021. Critical review on hazardous pollutants in water environment: Occurrence, monitoring, fate, removal technologies and risk assessment. Science of The Total Environment 797, 149134. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.scitotenv.2021.149134\u003c/span\u003e\u003cspan address=\"10.1016/j.scitotenv.2021.149134\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSchmidt, G.H., Ibrahim, N.M.M., Abdallah, M.D., 1992. Long-term effects of heavy metals in food on developmental stages of Aiolopus thalassinus (Saltatoria: Acrididae). Archives of Environmental Contamination and Toxicology 23, 375\u0026ndash;382. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/BF00216248\u003c/span\u003e\u003cspan address=\"10.1007/BF00216248\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSildanchandra, W., Crane, M., 2000. Influence of sexual dimorphism in Chironomus riparius Meigen on toxic effects of cadmium. Environmental Toxicology and Chemistry 19, 2309\u0026ndash;2313. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1002/etc.5620190921\u003c/span\u003e\u003cspan address=\"10.1002/etc.5620190921\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSorvari, J., Rantala, L.M., Rantala, M.J., Hakkarainen, H., Eeva, T., 2007. Heavy metal pollution disturbs immune response in wild ant populations. Environmental Pollution 145, 324\u0026ndash;328. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.envpol.2006.03.004\u003c/span\u003e\u003cspan address=\"10.1016/j.envpol.2006.03.004\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSu, H., Wu, M., Yang, Yong, Deng, Y., Yang, Yi-zhong, Sun, Q., 2023. Tissue distribution of cadmium and its effect on reproduction in Spodoptera exigua. Journal of Integrative Agriculture. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.jia.2023.08.010\u003c/span\u003e\u003cspan address=\"10.1016/j.jia.2023.08.010\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTabassum, S., Kotnala, C.B., Salman, M., Tariq, M., Khan, A.H., Khan, N.A., 2024. The impact of heavy metal concentrations on aquatic insect populations in the Asan Wetland of Dehradun, Uttarakhand. Scientific Reports 14, 4824. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1038/s41598-024-52522-5\u003c/span\u003e\u003cspan address=\"10.1038/s41598-024-52522-5\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003evan Capelleveen, H.E., van Straalen, N.M., van den Berg, M., van Wachem, E. (Eds.), 1986. Avoidance as a mechanism of tolerance for lead in terrestrial arthropods. Nederlandse Entomologische Vereniging, Amsterdam, The Netherlands.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003evan Straalen, N.M., Hoffmann, A.A., 2000. Review of experimental evidence for physiological costs of tolerance to toxicants., in: Demography in Ecotoxicology. John Wiley and Sons, pp. 147\u0026ndash;161.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVandenberg, L.N., Colborn, T., Hayes, T.B., Heindel, J.J., Jacobs, D.R., Jr., Lee, D.H., Shioda, T., Soto, A.M., vom Saal, F.S., Welshons, W.V., Zoeller, R.T., Myers, J.P., 2012. Hormones and endocrine-disrupting chemicals: low-dose effects and nonmonotonic dose responses. Endocrine reviews 33, 378\u0026ndash;455. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1210/er.2011-1050\u003c/span\u003e\u003cspan address=\"10.1210/er.2011-1050\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVardhan, K.H., Kumar, P.S., Panda, R.C., 2019. A review on heavy metal pollution, toxicity and remedial measures: Current trends and future perspectives. Journal of Molecular Liquids 290, 111197. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.molliq.2019.111197\u003c/span\u003e\u003cspan address=\"10.1016/j.molliq.2019.111197\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVerma, N., Rachamalla, M., Kumar, P.S., Dua, K., 2023. Chapter 6 - Assessment and impact of metal toxicity on wildlife and human health, in: Shukla, S.K., Kumar, S., Madhav, S., Mishra, P.K. (Eds.), Metals in Water. Elsevier, pp. 93\u0026ndash;110. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/B978-0-323-95919-3.00002-1\u003c/span\u003e\u003cspan address=\"10.1016/B978-0-323-95919-3.00002-1\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVlahović, M., Ilijin, L., Lazarević, J., Mrdaković, M., Gavrilović, A., Matić, D., Mataruga, V.P., 2014. Cadmium-induced changes of gypsy moth larval mass and protease activity. Comparative Biochemistry and Physiology Part C: Toxicology \u0026amp; Pharmacology 160, 9\u0026ndash;14. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.cbpc.2013.11.002\u003c/span\u003e\u003cspan address=\"10.1016/j.cbpc.2013.11.002\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVlahović, M., Matić, D., Mutić, J., Trifković, J., Đurđić, S., Perić Mataruga, V., 2017. Influence of dietary cadmium exposure on fitness traits and its accumulation (with an overview on trace elements) in Lymantria dispar larvae. Comparative Biochemistry and Physiology Part C: Toxicology \u0026amp; Pharmacology 200, 27\u0026ndash;33. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.cbpc.2017.06.003\u003c/span\u003e\u003cspan address=\"10.1016/j.cbpc.2017.06.003\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWasserstein, R.L., Lazar, N.A., 2016. The ASA Statement on p-Values: Context, Process, and Purpose. The American Statistician 70, 129\u0026ndash;133. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1080/00031305.2016.1154108\u003c/span\u003e\u003cspan address=\"10.1080/00031305.2016.1154108\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWu, G.-X., Ye, G.-Y., Hu, C., Cheng, J.-A., 2006. Accumulation of cadmium and its effects on growth, development and hemolymph biochemical compositions in Boettcherisca peregrina larvae (Diptera: Sarcophagidae). Insect Science 13, 31\u0026ndash;39. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/j.1744-7917.2006.00065.x\u003c/span\u003e\u003cspan address=\"10.1111/j.1744-7917.2006.00065.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZhang, Y., Wolosker, M.B., Zhao, Y., Ren, H., Lemos, B., 2020. Exposure to microplastics cause gut damage, locomotor dysfunction, epigenetic silencing, and aggravate cadmium (Cd) toxicity in Drosophila. Science of The Total Environment 744, 140979. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.scitotenv.2020.140979\u003c/span\u003e\u003cspan address=\"10.1016/j.scitotenv.2020.140979\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZulfiqar, U., Jiang, W., Xiukang, W., Hussain, S., Ahmad, M., Maqsood, M.F., Ali, N., Ishfaq, M., Kaleem, M., Haider, F.U., Farooq, N., Naveed, M., Kucerik, J., Brtnicky, M., Mustafa, A., 2022. Cadmium phytotoxicity, tolerance, and advanced remediation approaches in agricultural soils; A comprehensive review. Frontiers in Plant Science 13. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3389/fpls.2022.773815\u003c/span\u003e\u003cspan address=\"10.3389/fpls.2022.773815\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Cadmium, Heavy metal, Moth, Growth, Bioaccumulation, Insect","lastPublishedDoi":"10.21203/rs.3.rs-5439205/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5439205/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eOur environment is increasingly contaminated by various substances, mainly from industrial and agricultural activities. Among the main pollutants, heavy metals, are highly toxic to a wide range of organisms including invertebrates. In our study, we focused on the impacts of cadmium on the model pest moth \u003cem\u003eSpodoptera littoralis\u003c/em\u003e. In laboratory conditions, larvae were exposed to a diet contaminated with nine different cadmium concentrations ranging from 7.80 \u0026micro;g/g to 50 mg/g of food, over six developmental stages. Our results show that sublethal doses of cadmium have a significant effect on food consumption and relative larval growth rate over a 6-day period, and to a lesser extent, affect larval mass at higher doses. Adverse effects were noted at a sublethal dose of 31.25 \u0026micro;g/g, with particularly severe outcomes at the highest lethal doses ranging from 0.5 to 50 mg/g, where 100% mortality was observed after treatment. While higher doses from 250 \u0026micro;g/g influenced larval mass, most larvae recovered and regained mass, except for those exposed to the two highest doses just prior metamorphosis. In addition, our results indicated that cadmium accumulation reached its maximum at pupal stage and decreased in adults, suggesting that efficient detoxification processes occur during metamorphosis, with significant amounts of cadmium that were excreted through the integument and exuviae at the pre-pupal stage. Moreover, moths that ingested cadmium at larval stage have stored most of it in their digestive tract, from where it has been transferred to various parts of the body, including the head and antennae, via haemolymph.\u003c/p\u003e","manuscriptTitle":"Dose-dependent effects of cadmium on food consumption, mass, and bioaccumulation in the moth Spodoptera littoralis","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-12-02 13:41:22","doi":"10.21203/rs.3.rs-5439205/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"eabfe659-9635-4de8-b3a2-349e57b71b01","owner":[],"postedDate":"December 2nd, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-03-18T14:53:53+00:00","versionOfRecord":[],"versionCreatedAt":"2024-12-02 13:41:22","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-5439205","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5439205","identity":"rs-5439205","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}